Methods of treating kidney diseases

ABSTRACT

Provided herein are methods of treating kidney diseases in a subject in need thereof.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of PCT/US2020/065311, filed on Dec.16, 2020, which claims priority to U.S. Provisional Application No.62/949,115, filed on Dec. 17, 2019, U.S. Provisional Application No.63/005,003, filed on Apr. 3, 2020, U.S. Provisional Application No.63/072,699, filed Aug. 31, 2020, U.S. Provisional Application No.63/084,739, filed on Sep. 29, 2020 and U.S. Provisional Application No.63/125,205, filed on Dec. 14, 2020, which are incorporated by referencein their entirety.

BACKGROUND

IgA nephropathy (IgAN) is the most common primary glomerulonephritisworldwide. Aberrant glycosylation of IgA1 results in increased serumlevels of galactose-deficient IgA1 (Gd-IgA1) that are recognized byglycan-specific IgA and IgG autoantibodies. Aggregates of the immunecomplexes are formed in situ and/or deposited in the glomerularmesangium. This promotes proliferation of mesangial cells, increasedsynthesis of extracellular matrix proteins, cytokines, chemokines, andinfiltration of immune cells into the surrounding tissue. Accordingly,disease progression involves (1) production of Gd-IgA1; and (2) itsrecognition by antiglycan autoantibodies; which (3) form immunecomplexes in the kidney; and (4) activate mesangial cells. See, e.g.,Penfold et al., Int. J. Nephrol. and Renovascular Dis. 11, pp. 137-148(2017).

Unlike other progressive kidney diseases such as diabetic nephropathy,IgAN occurs primarily in subjects in their 20s and 30s who are otherwisehealthy. Patients present with a range of symptoms, typically includingmicro- or macro-hematuria and increased protein excretion in the urine.Patients may also present with hypertension as a result of sustainedrenal damage. Current therapeutic approaches merely provide supportivecare, including administration of the maximum tolerable dose of anangiotensin converting enzyme inhibitor or angiotensin-receptor blocker,or administration of immunosuppressive drugs, whose benefits are largelyoutweighed by adverse reactions. Ultimately, 30-40% of patients willdevelop end-stage renal disease (ESRD) within 20-30 years of diagnosisof IgAN. In the interim, patients experience numerous symptoms thatsignificantly degrade their quality of life, in addition to decliningrenal function. Patients with IgAN often exhibit significantly increasedexpressions of endothelin 1 (ET-1) and ET-RA in the kidney. Increasedexpression of endothelins positively correlates with proteinuria, one ofthe hallmark symptoms of IgAN.

SUMMARY

Atrasentan is a selective endothelin A (ETA) receptor antagonist (ETA Ki˜34 pM; ETB Ki ˜63 nM, ETA selectivity ˜1800x). See, e.g. Wu-Wong etal., Clin. Sci. (Lond.), 103(48), pp. 107s-111s (2002). Selective ETAreceptor antagonists block ETA function, while minimally effecting theETB receptor, providing beneficial renal effects including vasodilationand reduction of inflammation, while still enabling ET-1 clearance. Seee.g., Jandeleit-Dahm and Watson, Curr. Opin. Nephrol. Hypertens., 21(1),pp. 66-71 (2012); see also, Nakamura, et al., Nephron, Vol. 72, pp.454-460 (1996). While ETA receptor antagonists increase sodium and waterretention by the kidney, this is typically clinically manageable. See,e.g., Saleh, et al., J. Pharm. Exp. Ther., 338(1), pp. 263-270 (2011).Atrasentan has been shown to be effective in patients with diabetickidney disease (DKD), significantly reducing the risk of renal eventsdefined as a doubling of serum creatinine or end-stage kidney disease.See, e.g., Heerspink, et al., The Lancet, 393, pp 1937-1947 (2019). DKDis considered a secondary glomerular disease, where kidney diseasedevelops secondarily to an identified systemic cause, in the case of DKDas a microvascular complication to long standing diabetes. See, e.g.Dattani and McAdoo, Medicine, 47(10), pp. 644-648 (2019). Thepathogenesis of DKD is multifactorial and complex. Chronically elevatedblood glucose levels due to diabetes which causes glucose toxicity torenal cells, especially kidney endothelial cells, and systemic and renalhemodynamic factors associated with hypertension that result in shearstress being transmitted to resident glomerular cells are the keypathogenic drivers of DKD. See, e.g. Thomas et al., Nat. Rev. DiseasePrimers. 1, pp. 15018-15026 (2015). Multiple factors dysregulated in thediabetic milieu, including metabolic components such as hyperglycemia,dyslipidemia and oxidative stress, and hemodynamic factors such asvasoactive substances associated with hypertension, all stimulate renalET-1 formation. In addition, DKD is typically observed in olderpopulations due to the requirement for long-standing diabetes prior tothe manifestation of DKD, and aging is also associated with increasedET-1 production in the kidney. See, e.g. Kohan, Kidney Int., 86(5), pp.896-904 (2014). Combined, this all provides a sound scientific rationalefor the treatment of DKD with the ETA receptor blocker atrasentan. SeeDhaun, et al., Hypertension, Vol. 57, pp. 772-779 (2011).

In contrast to DKD, IgA nephropathy is considered a primary glomerulardisease where localized or intrinsic renal pathology is present. Thepeak incidence of IgA nephropathy is in young individuals in theirsecond or third decade of life and is a disease that is auto-immune inorigin, unlike DKD. IgA nephropathy results from pathogenic IgA/immunecomplex deposition in the glomerular mesangium. See, e.g., Lai, et al.,Nature Reviews Disease Primers, 2, pp. 16001, 2016. Definitive diagnosisrequires kidney biopsy and demonstration of mesangial IgA deposition byimmunofluorescence microscopy. Recent advances in the understanding ofthe initiating events triggering IgA nephropathy have revealed that anaberrant mucosal immune response stimulates the production of galactosedeficient IgA1 which is recognized as an autoantigen by circulatingantiglycan autoantibodies. Immune recognition results in the formationof nephritogenic immune complexes that deposit in the kidney andactivate mesangial cells. Activated mesangial cells proliferate andproduce excess amounts of extracellular matrix components, cytokines andchemokines. See, e.g., Suzuki, et al., J. Am. Soc. Nephrol., Vol. 22,pp. 1795-1803 (2011). Up to 40% of patients with biopsy-proven IgAnephropathy progress to end stage kidney disease at some point duringlong term follow up. The role of ET-1 or the ETA receptor in mesangialcell activation has not been previously reported and atrasentan has notbeen previously tested in IgA nephropathy. As further described herein,atrasentan can be administered at an efficacious dose with acceptabletoxicity, and has the appropriate selectivity to minimize undesired sideeffects while still treating the underlying IgAN and improving subjects'quality of life. Some embodiments provide a method of inhibitingmesangial cell activation in a subject having IgA nephropathy,comprising administering a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof, to thesubject; wherein the subject has not been previously diagnosed with oneor more of diabetic nephropathy, HIV/AIDS, or acute kidney failure.

Some embodiments provide a method of inhibiting PDGF signaling (e.g.,decreasing the activity and/or expression of one or more of PIK3R1,PDGFRA, NFKBIA, PIK3CG, PLA2G4A, TIAM1, PDGFB, NFKB1, and MAP3K1) in amesangial cell in a subject having IgA nephropathy, comprisingadministering a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof, to the subject; wherein thesubject has not been previously diagnosed with one or more of diabeticnephropathy, HIV/AIDS, or acute kidney failure.

Some embodiments provide a method of treating IgA nephropathy,comprising administering a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof, to a subjectin need thereof; wherein the subject has not been previously diagnosedwith one or more of diabetic nephropathy, HIV/AIDS, or acute kidneyfailure.

Some embodiments provide a method of treating IgA nephropathy,comprising administering a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof, to a subjectin need thereof; wherein the subject does not have one or more ofdiabetic nephropathy, HIV/AIDS, or acute kidney failure.

Some embodiments provide a method of treating IgA nephropathy,comprising administering a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof, to a subjectin need thereof; wherein the subject does not suffer from one or more ofdiabetic nephropathy, HIV/AIDS, or acute kidney failure.

Some embodiments provide a method of treating IgA nephropathy,comprising administering a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof, to a subjectin need thereof; wherein the subject has not been previously diagnosedwith one or more of diabetic nephropathy, HIV/AIDS, prostate cancer, oracute kidney failure.

Some embodiments provide a method of treating IgA nephropathy,comprising administering a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof, to a subjectin need thereof; wherein the subject has not been previously diagnosedwith one or more of diabetic nephropathy, HIV-related nephropathy,prostate cancer, or acute kidney failure.

Some embodiments provide a method of treating IgA nephropathy,comprising administering a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof, to a subjectin need thereof; wherein the subject has not been previously diagnosedwith one or more of diabetic nephropathy, HIV-related nephropathy, oracute kidney failure.

Some embodiments provide a method of treating IgA nephropathy,comprising administering a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof, to a subjectin need thereof; wherein the subject is not being treated for one ormore of diabetic nephropathy, HIV-related nephropathy, or acute kidneyfailure.

Some embodiments provide a method of treating IgA nephropathy,comprising administering a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof, to a subjectin need thereof; wherein the subject has been determined to havecontrolled serum glucose levels; and wherein the subject has not beendiagnosed with one or more of HIV-related nephropathy or acute kidneyfailure.

Some embodiments provide a method of decreasing renal inflammationand/or fibrosis in a subject having IgA nephropathy, comprisingadministering a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof, to a subject in need thereof.

Some embodiments provide a method of decreasing the occurrence ofhematuria in a subject having IgA nephropathy, comprising administeringa therapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, to a subject in need thereof.

Some embodiments provide a method of stabilizing eGFR in a subjecthaving IgA nephropathy, comprising administering a therapeuticallyeffective amount of atrasentan, or a pharmaceutically acceptable saltthereof, to a subject in need thereof.

Some embodiments provide a method of decreasing the number ofIgA-nephropathy associated disease flares in a subject having IgAnephropathy, comprising administering a therapeutically effective amountof atrasentan, or a pharmaceutically acceptable salt thereof, to asubject in need thereof.

Some embodiments provide a method of delaying the onset of ESRD in asubject having IgA nephropathy, comprising administering atherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, to a subject in need thereof.

Some embodiments provide a method of decreasing proteinuria in a subjecthaving IgA nephropathy, comprising administering a therapeuticallyeffective amount of atrasentan, or a pharmaceutically acceptable saltthereof, to a subject in need thereof.

Some embodiments provide a method of decreasing fatigue in a subjecthaving IgA nephropathy, comprising administering a therapeuticallyeffective amount of atrasentan, or a pharmaceutically acceptable saltthereof, to a subject in need thereof; wherein the subject has beendetermined not to suffer from one or more of diabetic nephropathy,HIV-related nephropathy, prostate cancer, or acute kidney failure.

Some embodiments provide a method of inhibiting mesangial cellactivation, comprising contacting a mesangial cell with an effectiveamount of atrasentan, or a pharmaceutically acceptable salt thereof.

Some embodiments provide a method of reducing activation of a mesangialcell in contact with an IgA immune complex, comprising contacting amesangial cell with an effective amount of atrasentan, or apharmaceutically acceptable salt thereof.

Some embodiments provide a method of treating IgA nephropathy in asubject in need thereof, comprising: a) determining that the subject haselevated serum Gd-IgA1 levels; and b) administering a therapeuticallyeffective amount of atrasentan, or a pharmaceutically acceptable saltthereof, to the subject

Some embodiments provide a method of treating IgA nephropathy in asubject in need thereof, comprising: a) determining that the subject haselevated levels of mesangial activation; and b) administering atherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, to the subject.

Some embodiments provide a method of treating IgA nephropathy in asubject in need thereof, comprising: a) determining that the subject haselevated levels of IgA-immune complexes in the kidney; and b)administering a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof, to the subject.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1A illustrates that treatment with atrasentan reduces theproliferation of primary human mesangial cells exposed to immunecomplexes derived from subjects with IgAN.

FIG. 1B illustrates the content of galactose-deficient (Gd)-IgA andtotal IgA following purification of immune complexes from IgAN orhealthy control serum.

FIG. 2A illustrates the decrease in proliferation (after 48 hours) ofprimary human mesangial cells stimulated with endothelin 1 (ET-1) upontreatment with increasing concentrations of atrasentan shown in logscale, demonstrating an IC₅₀ of about 5.1 nM.

FIG. 2B illustrates the decrease in proliferation (after 72 hours) ofprimary human mesangial cells stimulated with endothelin 1 (ET-1) upontreatment with increasing concentrations of atrasentan shown in logscale, demonstrating an IC₅₀ of about 50.8 nM.

FIG. 3A illustrates the increase in IL-6 production by primary humanmesangial cells exposed to ET-1 over 48 hours (about a 6-fold increase)and the decrease in IL-6 levels upon treatment with increasingconcentrations of atrasentan shown in log scale, demonstrating an IC₅₀of about 1 nM.

FIG. 3B illustrates the increase in IL-6 production by primary humanmesangial cells exposed to ET-1 over 72 hours (about a 2.2-foldincrease) and the decrease in IL-6 levels upon treatment with increasingconcentrations of atrasentan shown in log scale, demonstrating an IC₅₀of about 1 nM.

FIG. 4 illustrates UACR levels in g-ddY mice, at baseline, prior toatrasentan administration, and following approximately 5 days oftreatment with atrasentan at 0 (control), 10, 20 or 30 mg/kg/day in thedrinking water. (*P<0.05 compared to baseline levels, paired t-test).

FIG. 5 illustrates the change in UACR levels (% of baseline) in g-ddYmice, following approximately 5 days of treatment with atrasentan at 0(control), 10, 20 or 30 mg/kg/day in the drinking water. (*P<0.05compared to control group (0 mg/kg/day), unpaired t-test).

FIG. 6 illustrates the top 100 differentially expressed genes in kidneytissue following treatment of g-ddY mice with atrasentan at 0 (control),10, 20, or 30 mg/kg/day. Heatmap shows row-wise z-transformed counts permillion (CPM) values for each of the top 100 differentially expressedgenes.

FIG. 7A illustrates differential gene expression in kidney tissuefollowing treatment of g-ddY mice with atrasentan at 0 (control) and 10mg/kg/day. Black dots indicate genes not significantly differentiallyexpressed. Gray dots above 0 log-fold-change indicate upregulated genes.Gray dots below 0 log-fold change indicate downregulated genes.

FIG. 7B illustrates differential gene expression in kidney tissuefollowing treatment of g-ddY mice with atrasentan at 0 (control) and 20mg/kg/day. Black dots indicate genes not significantly differentiallyexpressed. Gray dots above 0 log-fold-change indicate upregulated genes.Gray dots below 0 log-fold change indicate downregulated genes.

FIG. 7C illustrates differential gene expression in kidney tissuefollowing treatment of g-ddY mice with atrasentan at 0 (control) and 30mg/kg/day. Black dots indicate genes not significantly differentiallyexpressed. Gray dots above 0 log-fold-change indicate upregulated genes.Gray dots below 0 log-fold change indicate downregulated genes.

FIG. 8A illustrates differentially expressed genes in kidney tissue in asubset of genes associated with an ET1 gene signature followingtreatment of g-ddY mice with atrasentan at 0 (control) and 10 mg/kg/day.

FIG. 8B illustrates differentially expressed genes in kidney tissue in asubset of genes associated with an ET1 gene signature followingtreatment of g-ddY mice with atrasentan at 0 (control) and 20 mg/kg/day.

FIG. 8C illustrates differentially expressed genes in kidney tissue fora subset of genes associated with an ET1 gene signature followingtreatment of g-ddY mice with atrasentan at 0 (control) and 30 mg/kg/day.

FIG. 9A illustrates the significantly enriched (FDR <0.05) hallmark genesets following treatment of g-ddY mice with atrasentan 10 mg/kg/daycompared to treatment of atrasentan at 0 mg/kg/day (control). Negativenormalized enrichment score (NES) indicates under-expression andpositive NES indicates over-expression.

FIG. 9B illustrates the significantly enriched (FDR <0.05) hallmark genesets following treatment of g-ddY mice with atrasentan 20 mg/kg/daycompared to treatment of atrasentan at 0 mg/kg/day (control). Negativenormalized enrichment score (NES) indicates under-expression andpositive NES indicates over-expression.

FIG. 9C illustrates the significantly enriched (FDR <0.05) hallmark genesets following treatment of g-ddY mice with atrasentan 30 mg/kg/daycompared to treatment of atrasentan at 0 mg/kg/day (control). Negativenormalized enrichment score (NES) indicates under-expression andpositive NES indicates over-expression.

FIG. 10 illustrates the concordance of significantly enriched (FDR<0.05) hallmark gene set pathways following treatment of g-ddY mice with10, 20 or 30 mg/kg/day compared to atrasentan at 0 (control), followingtreatment of ET-1 stimulated HRMC with atrasentan at 1 nM and 25 nM, andET-1 stimulated HRMC compared to untreated HRMC (control).

FIG. 11A illustrates the overlap in gene signature between publishedhuman mesangial single cell signature (Lake et al., A single-nucleusRNA-sequencing pipeline to decipher the molecular anatomy andpathophysiology of human kidneys. Nat. Comm., 10(1), 1-15 (2018);labeled as mesangial), differentially expressed genes followingtreatment of g-ddY mice with atrasentan at 30 mg/kg/day (labeled asg-ddY), and differentially expressed genes following treatment of HRMCwith atrasentan at 25 nM in the presence of ET1 (labeled as HRMC).

FIG. 11B illustrates the expression levels (row wise z-transformedvalues of log 2 counts per million) of 44 genes overlapped between humanmesangial single cell signature and differentially expressed genesfollowing treatment of g-ddY mice with atrasentan at 30 mg/kg/day.

FIG. 12A illustrates a comparison between potential biomarkers based onhuman and mouse with an “x” indicating translatability and whether thepotential biomarker is in plasma/serum and/or urine. Notably, each ofthe relevant markers are found in both humans and mice (e.g., g-ddYmice).

FIG. 12B illustrates log 2(CPM) for each potential biomarker followingtreatment of g-ddY mice with atrasentan at 0 (control), 10, 20 or 30mg/kg/day.

Table 1 Top 40 DEGs (25 upregulated and 15 downregulated) in treatmentof g-ddY mice with atrasentan at 10 mg/kg/day compared to atrasentan at0 mg/kg/day (control).

Table 2 Top 50 DEGs (25 upregulated and 25 downregulated) in treatmentof g-ddY mice with atrasentan at 20 mg/kg/day compared to atrasentan at0 mg/kg/day (control).

Table 3 Top 50 DEGs (25 upregulated and 25 downregulated) in treatmentof g-ddY mice with atrasentan at 30 mg/kg/day compared to atrasentan at0 mg/kg/day (control).

Table 4 The enriched signaling pathways following treatment of g-ddYmice with atrasentan at 10, 20 or 30 mg/kg/day compared to atrasentan at0 mg/kg/day (control). Z-score values indicate the magnitude of theeffect seen at each dosage.

Table 5 Gene Set Enrichment Analysis showing the enrichment in hallmarkgene sets following treatment of g-ddY mice with atrasentan at 10, 20 or30 mg/kg/day compared to atrasentan at 0 mg/kg/day (control). NES is thenormalized enrichment score indicating the representation of DEGs in thegene set and accounted for gene set size. NES of <−1.5 or >1.5 isconsidered to be biologically significant. Adj p-value is the estimatedprobability that the calculated NES of the given enriched gene set is afalse positive result.

Table 6 list of upstream regulators grouped by cytokine and growthfactor molecular types. P-value of overlap indicates the significance ofenrichment based on the number of overlapping genes between the datasetand the gene targets in IPKB. Activation z-score threshold of <−2 or >2and p-value of overlap threshold of <0.05 was considered significant.

Table 7 List of upstream regulators grouped by transmembrane receptor,G-protein coupled receptor, and protein complexes molecular types.P-value of overlap indicates the significance of enrichment based on thenumber of overlapping genes between the dataset and the gene targets inIPKB. Activation z-score threshold of <−2 or >2 and p-value of overlapthreshold of <0.05 was considered significant.

Table 8 List of upstream regulators grouped by transcription regulatorand ligand-dependent nuclear receptor molecular types. P-value ofoverlap indicates the significance of enrichment based on the number ofoverlapping genes between the dataset and the gene targets in IPKB.Activation z-score threshold of <−2 or >2 and p-value of overlapthreshold of <0.05 was considered significant.

Table 9 Gene expression of the components of the NF-kB signalingpathways following treatment of g-ddY mice with atrasentan at 10, 20 or30 mg/kg/day compared to atrasentan at 0 mg/kg/day (control).

Table 10 Gene expression of the components of the IL6 signaling pathwaysfollowing treatment of g-ddY mice with atrasentan at 10, 20 or 30mg/kg/day compared to atrasentan at 0 mg/kg/day (control).

Table 11 Gene expression of the components of the PDGF signalingpathways following treatment of g-ddY mice with atrasentan at 10, 20 or30 mg/kg/day compared to atrasentan at 0 mg/kg/day (control).

Table 12 Gene expression of the components of the cell proliferationsignaling pathways (mitotic spindle and G2M cell cycle checkpoint)following treatment of g-ddY with atrasentan at 10, 20, or 30 mg/kg/daycompared to atrasentan at 0 mg/kg/day (control).

Table 13 Gene expression of the components of the inflammatory responsesignaling pathways following treatment of g-ddY with atrasentan at 10,20, or 30 mg/kg/day compared to atrasentan at 0 mg/kg/day (control).

Table 14 Gene expression of the 44 genes associated with mesangial cellsignature following treatment of g-ddY with atrasentan at 10, 20, or 30mg/kg/day compared to atrasentan at 0 mg/kg/day (control).

DETAILED DESCRIPTION

A. Definitions

In order that the present disclosure can be more readily understood,certain terms are first defined. As used in this application, except asotherwise expressly provided herein, each of the following terms shallhave the meaning set forth below. Additional definitions are set forththroughout the application.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this disclosure is related. For example, the ConciseDictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed.,2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd ed.,1999, Academic Press; and the Oxford Dictionary Of Biochemistry AndMolecular Biology, Revised, 2000, Oxford University Press, provide oneof skill with a general dictionary of many of the terms used in thisdisclosure. For purposes of the present disclosure, the following termsare defined.

Units, prefixes, and symbols are denoted in their Systéme Internationalde Unites (SI) accepted form. Numeric ranges are inclusive of thenumbers defining the range. The headings provided herein are notlimitations of the various aspects of the disclosure, which can be hadby reference to the specification as a whole. Accordingly, the termsdefined immediately below are more fully defined by reference to thespecification in its entirety.

The terms “a,” “an,” or “the” as used herein not only include aspectswith one member, but also include aspects with more than one member. Forinstance, the singular forms “a,” “an,” and “the” include pluralreferents unless the context clearly dictates otherwise. Thus, forexample, reference to “a cell” includes a plurality of such cells andreference to “the agent” includes reference to one or more agents knownto those skilled in the art, and so forth.

The term “or” as used herein should in general be construednon-exclusively. For example, a claim to “a composition comprising A orB” would typically present an aspect with a composition comprising bothA and B. “Or” should, however, be construed to exclude those aspectspresented that cannot be combined without contradiction (e.g., acomposition pH that is between 9 and 10 or between 7 and 8).

The group “A or B” is typically equivalent to the group “selected fromthe group consisting of A and B.”

The term “and/or” where used herein is to be taken as specificdisclosure of each of the two specified features or components with orwithout the other. Thus, the term “and/or” as used in a phrase such as“A and/or B” herein is intended to include “A and B,” “A or B,” “A”(alone), and “B” (alone). Likewise, the term “and/or” as used in aphrase such as “A, B, and/or C” is intended to encompass each of thefollowing aspects: A, B, and C; A, B, or C; A or C; A or B; B or C; Aand C; A and B; B and C; A (alone); B (alone); and C (alone).

The terms “about” and “approximately” as used herein shall generallymean an acceptable degree of error for the quantity measured given thenature or precision of the measurements. Typical, exemplary degrees oferror are within 20 percent (%), within 10%, or within 5% of a givenvalue or range of values. Any reference to “about X” specificallyindicates at least the values X, 0.95X, 0.96X, 0.97X, 0.98X, 0.99X,1.01X, 1.02X, 1.03X, 1.04X, and 1.05X. Thus, “about X” is intended toprovide written description support for a claim limitation of, e.g.,“0.98X.” The terms “about” and “approximately,” particularly inreference to a given quantity, encompass and describe the given quantityitself.

When “about” is applied to the beginning of a numerical range, itapplies to both ends of the range. Thus, “from about 5 to 20%” isequivalent to “from about 5% to about 20%.” When “about” is applied tothe first value of a set of values, it applies to all values in thatset. Thus, “about 0.5, 0.75, or 1.0 mg” is equivalent to “about 0.5,about 0.75, or about 1.0 mg.”

As used herein, the term “about”, when preceding a series of peakpositions for X-ray powder diffraction (e.g., 20 values), means that allof the peaks of the group which it precedes are reported in terms ofangular positions with a variability of ±0.1°. Accordingly, for example,the phrase about 8.3°, 9.7°, 10.0°, 13.0°, 15.6°, 17.2° or 19.5° means8.3°±0.1°, 9.7°±0.1°, 10.0°±0.1°, 13.0°±0.1°, 15.6°±0.1°, 17.2°±0.1°, or19.5°+0.1°.

“Treatment” or “therapy” of a subject refers to any type of interventionor process performed on, or the administration of an active agent to,the subject with the objective of reversing, alleviating, ameliorating,inhibiting, or slowing down, the onset, progression, development,severity, or recurrence of a symptom, complication, condition, orbiochemical indicia associated with a disease.

“Administering” or “administration” refer to the physical introductionof a therapeutic agent to a subject, using any of the various methodsand delivery systems known to those skilled in the art. Routes ofadministration can include oral, intravenous, intramuscular,subcutaneous, intraperitoneal, spinal or other parenteral routes ofadministration, for example by injection or infusion (e.g., intravenousinfusion). Administration can also be performed, for example, once, aplurality of times, and/or over one or more extended periods.

The term “prophylactic” or “prophylactically” refers to any type ofintervention or process performed on, or the administration of an activeagent to, the subject with the objective of protecting or preventing adisease or condition from developing or at least not developing fully(e.g., to reduce the symptoms or severity of the disease or condition)such as in the development of a side effect.

A “subject” includes any human or non-human animal. The term “non-humananimal” includes, but is not limited to, vertebrates such as non-humanprimates, sheep, dogs, and rodents such as mice, rats, and guinea pigs.In some embodiments, the subject is a human. The terms “subject” and“patient” and “individual” are used interchangeably herein.

An “effective amount” or “therapeutically effective amount” or“therapeutically effective dosage” of a drug or therapeutic agent is anyamount of the drug that, when used alone or in combination with anothertherapeutic agent, slowing down the onset of a disease or promotesdisease regression evidenced by a decrease in severity of diseasesymptoms, an increase in frequency and duration of disease symptom-freeperiods, or a ameliorating an impairment or disability due to thedisease affliction. The ability of a therapeutic agent to promotedisease regression can be evaluated using a variety of methods known tothe skilled practitioner, such as in human subjects during clinicaltrials, in animal model systems predictive of efficacy in humans, or byassaying the activity of the agent in in vitro assays.

The phrase “pharmaceutically acceptable” indicates that the substance orcomposition must be compatible chemically and/or toxicologically, withthe other ingredients comprising a formulation, and/or the mammal beingtreated therewith.

As used herein, “polymorphs” refer to distinct solids sharing the samemolecular formula, yet each polymorph may have distinct solid statephysical properties. A single compound may give rise to a variety ofpolymorphic forms where each form has different and distinct solid statephysical properties, such as different solubility profiles, meltingpoint temperatures, flowability, dissolution rates and/or differentX-ray diffraction peaks. These practical physical characteristics areinfluenced by the conformation and orientation of molecules in the unitcell, which defines a particular polymorphic form of a substance.Polymorphic forms of a compound can be distinguished in a laboratory byX-ray diffraction spectroscopy, such as X-ray powder diffraction(“XRPD”), and by other methods, such as infrared spectrometry.Additionally, polymorphic forms of the same drug substance or activepharmaceutical ingredient can be administered by itself or formulated asa drug product (pharmaceutical composition) and are well known in thepharmaceutical art to affect, for example, the solubility, stability,flowability, tractability and compressibility of drug substances and thesafety and efficacy of drug products. For more, see Hilfiker, Rolf(ed.), Polymorphism in the Pharmaceutical Industry. Weinheim, Germany:Wiley-VCH 2006.

As used herein, the term “amorphous” means a solid in a solid state thatis a non-crystalline state. Amorphous solids generally possesscrystal-like short range molecular arrangement, but no long range orderof molecular packing as found in crystalline solids. The solid stateform of a solid may be determined by polarized light microscopy, X-raypowder diffraction (“XRPD”), differential scanning calorimetry (“DSC”),or other standard techniques known to those of skill in the art.

As used herein, the term “crystalline” means a solid in a solid statehaving a regularly repeating arrangement of molecules or external faceplanes. The solid state form of a solid may be determined by polarizedlight microscopy, X-ray powder diffraction (“XRPD”), differentialscanning calorimetry (“DSC”), or other standard techniques known tothose of skill in the art. Accordingly, the term “crystalline purity,”as used herein, means the percentage of a certain crystalline polymorphof atrasentan or pharmaceutically acceptable salt thereof in a samplethat may contain amorphous atrasentan or a pharmaceutically acceptablesalt thereof, one or more additional crystalline polymorphs ofatrasentan or a pharmaceutically acceptable salt thereof, or mixturesthereof. When a crystalline polymorph of atrasentan or apharmaceutically acceptable salt thereof is described as having“substantial crystalline purity”, it means the polymorph issubstantially free (e.g., contains <10%, <5%, <2%, <1%, <0.5%, <0.1%, or<0.05%) of other polymorphs (amorphous and/or crystalline).

The term “chemical purity,” as used herein, means percentage of aparticular compound (e.g., atrasentan or a pharmaceutically acceptablesalt thereof) in a sample. Accordingly, atrasentan or a pharmaceuticallyacceptable salt thereof and compositions comprising or made therefrommay contain one or more impurity, including but not limited to: water,ethyl acetate, ethanol,(2R,3R,4S)-2-(4-methoxyphenyl)-4-(1,3-benzodioxol-5-yl)-1-(N-(n-butyl)aminocarbonylmethyl)pyrrolidine-3-carboxylicacid,(2R,3R,4S)-2-(4-methoxyphenyl)-4-(1,3-benzodioxol-5-yl)-1-((N-(n-butyl)-N-ethyl)aminocarbonylmethyl)pyrrolidine-3-carboxylicacid,(2R,4S)-2-(4-methoxyphenyl)-4-(1,3-benzodioxol-5-yl)-1-(N,N-di(n-butyl)aminocarbonylmethyl)pyrrolidine,or ethyl(2R,3R,4S)-2-(4-methoxyphenyl)-4-(1,3-benzodioxol-5-yl)-1-(N,N-di(n-butyl)aminocarbonylmethyl)pyrrolidine-3-carboxylate.When a sample of atrasentan or a pharmaceutically acceptable saltthereof is described as having “substantial purity”, the sample issubstantially free of impurities (e.g., contains <10%, <5%, <2%, <1%,<0.5%, <0.1%, or <0.05%).

The term “diastereomeric excess,” as used herein, means the amount ofone diastereomer of a compound (e.g., atrasentan or a pharmaceuticallyacceptable salt thereof) in a mixture which may have other diastereomersof the same compound in the mixture. The term “substantialdiastereomeric purity,” as used herein, means diastereomeric excessgreater than about 90%, 95%, 99%, 99.5%, 99.9%, or 100%.

As used herein, the term “pharmaceutically acceptable carrier” refers toa substance that aids the administration of an active agent to a cell,an organism, or a subject. “Pharmaceutically acceptable carrier” refersto a carrier or excipient that can be included in the compositions ofthe disclosure and that causes no significant adverse toxicologicaleffect on the subject. Non-limiting examples of pharmaceuticallyacceptable carriers include water, NaCl, normal saline solutions,lactated Ringer's, normal sucrose, normal glucose, binders, fillers,disintegrants, lubricants, coatings, sweeteners, flavors and colors,liposomes, dispersion media, microcapsules, cationic lipid carriers,isotonic and absorption delaying agents, and the like. The carrier mayalso be substances for providing the formulation with stability,sterility and isotonicity (e.g., antimicrobial preservatives,antioxidants, chelating agents and buffers), for preventing the actionof microorganisms (e.g. antimicrobial and antifungal agents, such asparabens, chlorobutanol, phenol, sorbic acid and the like) or forproviding the formulation with an edible flavor etc. In some instances,the carrier is an agent that facilitates the delivery of a smallmolecule drug or antibody to a target cell or tissue. One of skill inthe art will recognize that other pharmaceutical carriers are useful inthe present disclosure.

The term “expression” as used herein refers the level of protein or mRNAin a mammalian cell.

The term “activity” as used herein refers to one or more activities of aprotein, such as binding or enzymatic activity (e.g., one or more ofphosphorylation, dephosphorylation, nuclear import, transcriptionalactivation, transcriptional repression, and/or binding activity to asubstrate or a binding partner).

The term “IL-6 signaling” as used herein means the expression and/oractivity of one or more proteins in a signaling pathway beginning withactivation of an IL-6 receptor and ending in gene expression.Non-limiting examples of proteins in a signaling pathway beginning withactivation of an IL-6 receptor and ending in gene expression include anIL-6 receptor, JAK, STAT3, PI3K, Akt/PKB, IKKs, IkBs, NF-kB, MAPK, Ras,Raf, MEK, and ERK.

The term “NF-kB signaling” as used herein means the expression and/oractivity of one or more of IKKα, IKKβ, IkB, and NF-kB, and/or one ormore genes upregulated by activity of NF-kB (e.g., one or more of TNF-α,IL-1, CAM, COX-2, and iNOS).

The term “PDGF signaling” as used herein means the expression and/oractivity of one or more of PDGF receptor, PKC, PI3K, Src, Ras, ERK1/2,Rho, Rac, Akt, mTOR, NAPDH oxidase, MAPK, and cPLA2.

The term “SGLT-2 inhibitor” as used herein refers to a compound thatinhibits the Sodium Glucose Co-Transporter-2 (SGLT-2). SGLT-2 inhibitorsdisrupt reabsorption of glucose by the kidneys and thus exert aglucose-lowering effect. By enhancing glucosuria, independently ofinsulin, SGLT-2 inhibitors have been shown to treat Type 2 diabetes andto improve cardiovascular outcomes. See Wright, 2001, Am. J. Physiol.Renal Physiol. 280:F10; and Scheen, 2018, Circ. Res. 122:1439. In someembodiments, the term “SGLT-2 inhibitor” refers to compounds whoseprimary effect is inhibition of SGLT-2, but is not limited to compoundsthat only inhibit SGLT-2, thus including compounds that have otheractivities in addition to SGLT-2 inhibition (e.g., SGLT-1 inhibition).

In some embodiments, SGLT-2 inhibitors include compounds of a class ofdrugs known as gliflozins. In some embodiments, SGLT-2 inhibitorsinclude compounds that are approved as SGLT-2 inhibitors by a regulatoryagency such as the FDA or EMA. Non-limiting examples of SGLT-2inhibitors include bexagliflozin, canagliflozin (INVOKANA®),dapagliflozin (FARXIGA®), empagliflozin (JARDIANCE®), ertugliflozin(STEGLATRO™), ipragliflozin (SUGLAT®), luseogliflozin (LUSEFI®),remogliflozin, sergliflozin, licogliflozin, sotagliflozin (ZYNQUISTA™),and tofogliflozin.

In some embodiments, the SGLT-2 inhibitors include, but are not limitedto dapagliflozin, canagliflozin, ipragliflozin, empaglifozin,bexagliflozin, licogliflozin, janagliflozin (XZP-5695), tofogliflozin,ertugliflozin, henagliflozin (SHR-3824), enavogliflozin (DWP-16001),TA-1887(3-(4-cyclopropylbenzyl)-4-fluoro-1-(β-D-glucopyranosyl)-1H-indole),indole-N-glycoside 18(3-(4-ethylbenzyl)-1-(β-D-glucopyranosyl)-1H-indole), sotagliflozin,luseogliflozin, sergliflozin etabonate (ethyl carbonate), remogliflozin,remogliflozin etabonate, and T-1095(((2R,3S,4S,5R,6S)-6-(2-(3-(benzofuran-5-yl)propanoyl)-3-hydroxy-5-methylphenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)etabonate).

In some embodiments, the SGLT-2 inhibitors include C-glycosides such asdapagliflozin, canagliflozin, ipragliflozin, empaglifozin,bexagliflozin, licogliflozin, janagliflozin (XZP-5695), tofogliflozin,ertugliflozin, henagliflozin (SHR-3824), enavogliflozin (DWP-16001). Insome embodiments, the SGLT-2 inhibitors include C-glycosides with abicyclic or spiro pyran group, such as tofogliflozin, ertugliflozin, andhenagliflozin (SHR-3824). In some embodiments, the SGLT-2 inhibitorsinclude C-glycosides that do not have a bicyclic or spiro pyran group,such as dapagliflozin, canagliflozin, ipragliflozin, empaglifozin,bexagliflozin, licogliflozin, janagliflozin (XZP-5695), andenavogliflozin (DWP-16001).

In some embodiments, the SGLT-2 inhibitors include N-glycosides such asTA-1887(3-(4-cyclopropylbenzyl)-4-fluoro-1-(β-D-glucopyranosyl)-1H-indole) andindole-N-glycoside 18(3-(4-ethylbenzyl)-1-(β-D-glucopyranosyl)-1H-indole).

In some embodiments, the SGLT-2 inhibitors include2-methylthio-C-glycosides, such as sotagliflozin.

In some embodiments, the SGLT-2 inhibitors includethiopyran-C-glycosides, such as luseogliflozin.

In some embodiments, the SGLT-2 inhibitors include 0-glycosides and0-glycoside prodrugs, such as sergliflozin etabonate (ethyl carbonate),remogliflozin, remogliflozin etabonate, and T-1095(((2R,3S,4S,5R,6S)-6-(2-(3-(benzofuran-5-yl)propanoyl)-3-hydroxy-5-methylphenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)etabonate).

In some embodiments, an SGLT-2 inhibitor, as defined herein, includesany compound exhibiting SGLT-2 inhibition activity. In some embodiments,an SGLT-2 inhibitor is selective for SGLT-2 over SGLT-1, for example, byhaving about 2-fold, about 5-fold, about 10-fold, about 20-fold, about50-fold, about 100-fold, about 200-fold, about 300-fold, about 400-fold,about 500-fold, about 750-fold, about 1,000-fold, about 1,250-fold,about 1,500-fold, about 1,750-fold, about 2,000-fold, about 2,500-fold,or any value in between, greater activity against SGLT-2 than againstSGLT-1. Exemplary SGLT-2 inhibitors can exhibit inhibition activity(IC₅₀) against SGLT-2 of less than about 1000 nM, less than about 500nM, less than about 200 nM, less than about 100 nM, less than about 50nM, less than about 25 nM, less than about 10 nM, or less than about 1nM as measured in an assay as described herein. In some embodiments,SGLT-2 inhibitors can exhibit inhibition activity (IC₅₀) against SGLT-2of less than about 25 nM, less than about 10 nM, less than about 5 nM,or less than about 1 nM as measured in an assay as provided herein. Anexemplary assay for determining SGLT-2 inhibitory activity is describedin Ryan, et al., Kidney International, Vol. 45, pp. 48-57 (1994).Briefly, CHO cells are stably transfected with cDNA encoding humanSGLT-2 (GenBank #M95549). Cells are washed and then incubated with 10 μM[¹⁴C]alpha-methyl glucopyranoside (AMG), and 10 μM inhibitor. The uptakeof [¹⁴C]AMG is quenched with cold buffer containing phlorizin, and cellsare lysed. Suitable reagents are then used to quantify the uptake of[¹⁴C]AMG.

SGLT-2 inhibitors include pharmaceutically acceptable salts, solvates,complexes, and salts of solvates thereof, for example, “dapagliflozin”includes salts of dapagliflozin (such as the hydrochloride salt) as wellas solvates (such as the propylene glycol hydrate); likewise,“canagliflozin” includes solvates (such as canagliflozin hemihydrate)and salts of solvates (such as the hydrochloride salt of the hydrate).Similarly, henagliflozin (SHR-3824) and dapagliflozin include complexes(such as the complexes henagliflozin proline and dapagliflozin proline,respectively).

As used herein, when a subject is described as having “controlled serumglucose levels”, it means the subject has a serum glucose level withinthe normal or healthy ranges. In some embodiments, the subject has afasting serum glucose level of between about 70 mg/dL and about 130mg/dL. For example, the subject has been determined to have a fastingserum glucose level of below about 130 mg/dL, 125 mg/dL, 120 mg/dL, 115mg/dL, 110 mg/dL, 105 mg/dL, 100 mg/dL, 95 mg/dL, 90 mg/dL, 85 mg/dL, 80mg/dL, or 75 mg/dL.

As used in the methods described herein, the term “reducing” refers to areduction in the indicated parameter relative to the baselinemeasurement (or measurements) of the same parameter in the subject takenprior to the initiation of administration with atrasentan or apharmaceutically acceptable salt thereof, or a reduction in theindicated parameter relative to the baseline measurement (ormeasurements) of the same parameter in a healthy subject (for example, asubject that does not have IgA nephropathy). Similarly, the term“increasing,” as used herein, refers to an increase in the indicatedparameter relative to the baseline measurement (or measurements) of thesame parameter in the subject taken prior to the initiation ofadministration with atrasentan or a pharmaceutically acceptable saltthereof, or an increase in the indicated parameter relative to thebaseline measurement (or measurements) of the same parameter in ahealthy subject (for example, a subject that does not have IgAnephropathy).

The term “glomerular filtration rate (GFR)” is defined as the volume offluid filtered from the renal (kidney) glomerular capillaries into theBowman's capsule per unit time. It is indicative of overall kidneyfunction. The glomerular filtration rate (GFR) can be calculated bymeasuring any chemical that has a steady level in the blood, and isfreely filtered but neither reabsorbed nor secreted by the kidneys. Therate therefore measured is the quantity of the substance in the urinethat originated from a calculable volume of blood. The GFR is typicallyrecorded in units of volume per time, e.g., milliliters per minute andthe formula below can be used: GFR=(Urine Concentration×UrineVolume)/Plasma Concentration. The GFR can be determined by injectinginulin into the plasma. Since inulin is neither reabsorbed nor secretedby the kidney after glomerular filtration, its rate of excretion isdirectly proportional to the rate of filtration of water and solutesacross the glomerular filter. A normal value is: GFR=90-125 mL/min/1.73m², in particular GFR=100-125 mL/min/1.73 m². Other principles todetermine GFR involve measuring 51Cr-EDTA, [125I]iothalamate or iohexol.The “estimated glomerular filtration rate (eGFR)” is defined as derivedat screening from serum creatinine values based on e.g., the ChronicKidney Disease Epidemiology Collaboration (CKD-EPI) equation, theCockcroft-Gault formula or the Modification of Diet in Renal Disease(MDRD) formula, which are all known in the art. “Stabilizing eGFR” asused herein means reducing the rate of decrease of eGFR and/orattenuating the rate of decline of eGFR. For example, the rate ofdecline of eGFR can be attenuated by at least about 20%; by at leastabout 30%; by at least about 40%; by at least about 50%; by at leastabout 60%; by at least about 70%; by at least about 80%; by at leastabout 90%; or by at least about 95%; or any value in between aftertreatment with atrasentan, or a pharmaceutically acceptable saltthereof. This attenuation can be after treatment, for example, for about1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks,about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10weeks, about 20 weeks, about 30 weeks, about 40 weeks, about 50 weeks,about 60 weeks, about 70 weeks, about 80 weeks, about 90 weeks, about100 weeks, about 110 weeks, about 120 weeks, about 130 weeks, about 140weeks, about 150 weeks, about 160 weeks, about 170 weeks, about 180weeks, about 190 weeks, or about 200 weeks, or any value in between. Insome embodiments, the subject has been treated with atrasentan or apharmaceutically acceptable salt thereof for between about 15 days andabout 30 days. In some embodiments, the subject has been treated withatrasentan or a pharmaceutically acceptable salt thereof for betweenabout 6 months and about 1 year.

“ESRD” is the abbreviation for end-stage renal disease. As used herein,the onset of ESRD is defined as the time point when the subject has aneGFR of below about 15 mL/min/1.73 m² and/or when the subject hasinitiated chronic dialysis. When a subject is defined to be “at a highrisk of progression to ESRD”, the subject has >1 g/day protein in theurine and/or eGFR <60 for at least about 3 months before the firstadministration of atrasentan or a pharmaceutically acceptable saltthereof.

“IgA-nephropathy associated disease flares” as used herein refer todisease flares associated with hematuria, worsening proteinuria,systemic manifestations, and declines in eGFR. Other symptoms associatedwith disease flares include: increased edema, fatigue, increasedhematuria, gross hematuria, and other symptoms which generallynegatively impact disease progression.

As used herein, when a subject is described to “maintain a potassiumlevel within the normal physiologic range”, the subject has a bloodpotassium level of from about 3.5 mEq/L to about 5.2 mEq/L.

As used herein, when a subject is described to “maintain a sodium levelwithin the normal physiologic range”, the subject has a blood sodiumlevel of from about 135 to about 145 mEq/L.

As used herein, the term “proteinuria” refers to the presence of proteinin the urine in excess of normal levels. “Proteinuria” includes“albuminuria” and “microalbuminuria”. Normal human levels of proteinappear in the urine in the range of about 0 to 30 mg/L, although for anygiven urine sample, the level may reach about 80 mg/L. For a 24 hoururine collection, normal human levels of urinary protein are in therange of about 0 to 150 mg. Proteinuria can be indicated by the ratio oftotal protein/creatinine in the urine (UPCR), or by the ration of aspecific protein, such as a urinary albumin/creatinine ratio (ACR) ofgreater than about 30 mg/g. Typically, the urinary UACR value in mg/gapproximately equals to the albumin excretion by the subject in mg/day.Proteinuria, including albuminuria and microalbuminuria, often leads toor is indicative of a disease, but is not limited to production of adisease. Proteinuria is intended to encompass all forms of proteinuria,including but not limited to physiological proteinuria; functionalproteinuria; and athletic proteinuria, which relates to a form offunctional proteinuria following excessive muscular exertion. Further,proteinuria covers benign proteinuria (also known as “essential”proteinuria), which refers to types or proteinuria that are not theresult of pathologic changes in the kidneys. Proteinuria also coverspathologic proteinuria, for example levels of protein in the urinegreater than normal physiological levels.

As used herein, the term “albuminuria” (also known as“macroalbuminuria”) refers to the presence of albumin in the urine inexcess of normal levels. Since urinary protein is predominantly albumin,normal human levels of urinary UACR are in the range of about 0 to 30mg/mmol. As used herein, the term “microalbuminuria” refers to thepresence of albumin in the urine, excreted at a rate of about 20 to 200μg/min or at a level of about 30 to 300 mg/L in humans. When defined bythe urinary ACR, “microalbuminuria” refers to a urinary UACR of greaterthan about 30 mg/g, or a urinary UACR of about 3.5 mg/mmol or greaterfor women and about 2.5 mg/mmol or greater for men. Microalbuminuria isoften an early warning of kidney disease, but can also be present forother reasons.

As used herein, the term “hematuria” refers to the presence of blood inthe urine. It may present as macroscopic hematuria (visible traces ofblood cells) or microscopic hematuria (microscopic traces of blood)within the urine. A confirmed indication of microhematuria is defined as3 or more red blood cells present per microscopic high-powered field(HPF) on a minimum of 3 properly collected urine samples. Microhematuriamay also be detected by urine dipstick (colorimetric comparisonestimate) at clinic. Hematuria (either microscopic or macroscopic) maybe asymptomatic (no additional symptoms associated with hematuria) orsymptomatic. Additional symptoms include dysuria (painful urination), afeeling of incomplete emptying of the bladder or increased frequency orurination, or flank pain.

“ALT” as used herein refers to alanine transaminase. “AST” as usedherein refers to aspartate transaminase.

The term “synergy” or “synergistic” is used herein to mean that theeffect of a combination of two or more therapeutic agents is greaterthan the sum of the effect of each agent when administered alone. See,e.g., Chou and Talalay, Advances in Enzyme Regulation (1984), 22, 27-55.A “synergistically effective amount” is an amount of the combination ofthe two or more therapeutic agents that results in a synergistic effect(as “synergistic” is defined herein). In some embodiments, asynergistically effective amount of a combination may be therapeuticallyeffective even when one or more of the compounds in the combination isadministered at a dose that would be sub-therapeutic when the compoundis administered alone.

It will be appreciated that different concentrations of each compoundmay be employed for various art-recognized factors, for example, thepatient's height, weight, sex, age and medical history. Exemplarysynergistic effects includes, but are not limited to, enhancedtherapeutic efficacy, decreased dosage at equal or increased level ofefficacy, reduced or delayed development of drug resistance, andsimultaneous enhancement or equal therapeutic actions (e.g., the sametherapeutic effect as at least one of the therapeutic agents) and areduction of unwanted drug effects (e.g. side effects and adverseevents) of at least one of the therapeutic agents.

In some embodiments, “synergistic effect” as used herein refers to acombination of atrasentan, or a pharmaceutically acceptable saltthereof, and one or more additional therapeutic agents (for example, aSGLT-2 inhibitor), producing an effect, for example, any of thebeneficial or desired results including clinical results as describedherein, which is greater than the sum of effect observed when theatrasentan, or a pharmaceutically acceptable salt thereof, and theSGLT-2 inhibitor are administered alone. Such clinical results include,but are not limited to treating IgA nephropathy, decreasing renalinflammation and/or fibrosis, decreasing hematuria, decreasingproteinuria (e.g., albuminuria), stabilizing eGFR, decreasing the numberof IgA-nephropathy associated disease flares, delaying the onset ofESRD, decreasing fatigue, reducing activation of a mesangial cell.

In some embodiments, “synergistic effect” as used herein refers to acombination of atrasentan, or a pharmaceutically acceptable salt thereofand a SGLT-2 inhibitor, providing a greater reduction in proteinuria,such as albuminuria, than the sum of effect observed when theatrasentan, or a pharmaceutically acceptable salt thereof, and theSGLT-2 inhibitor are administered alone.

In some embodiments, “synergistic effect” as used herein refers to acombination of atrasentan, or a pharmaceutically acceptable saltthereof, and an SGLT-2 inhibitor, producing a desired therapeutic effectand a reduction in the occurrence and/or severity of an unwanted drugeffect, side effect, or adverse event. In some embodiments, the unwanteddrug effect, side effect, or adverse event is associated with orobserved in monotherapy of atrasentan, or a pharmaceutically acceptablesalt thereof, or a SGLT-2 inhibitor. In some embodiments, the unwanteddrug effect, side effect, or adverse event is one or more of fluidretention, anemia, nausea, constipation, thirst, bone fractures,increased urination, urinary tract infection, yeast infection, vaginalitching, increased LDL cholesterol levels, increased brain natriureticpeptide (BNP) levels, acute sodium retention, and acute increases increatinine levels. In some embodiments, the fluid retention isassociated with a weight gain of greater than about 3 kg. In someembodiments, the increased BNP levels are greater than about 300 pg/mL.

As described herein, any concentration range, percentage range, ratiorange, or integer range is to be understood to include the value of anyinteger within the recited range and, when appropriate, fractionsthereof (such as one tenth and one hundredth of an integer), unlessotherwise indicated.

Unless otherwise stated, any reference to an amount of atrasentan inthis disclosure is based on the free equivalent weight of atrasentan.For example, 0.75 mg of atrasentan refers to 0.75 mg of atrasentan inthe free form or an equivalent amount of a salt form of atrasentan.

Various aspects of the disclosure are described in further detail in thefollowing subsections.

B. Introduction

Most subjects with IgAN first present with either single or episodicmacroscopic hematuria, or after detection of microscopic hematuriaand/or proteinuria during routine urine testing. In some cases, subjectspresent with acute kidney injury, such as a result of crescentic IgAN orgross hematuria causing tubular obstruction. Definitive diagnosis ofIgAN is typically is established by kidney biopsy, withimmunofluorescence and/or immunoperoxidase studies for IgA deposits.Prominent, globular deposits of IgA (sometimes accompanied by C3 andIgG) in the mesangium and less prominently along the glomerularcapillary wall are a hallmark of IgAN. Certain histopathologicalfeatures that correlate with long-term outcome include mesangialproliferation, endocapillary proliferation, segmental scarring, andtubular atrophy.

C. Methods of Treatment

In a normal and healthy human kidney, expressions of ET-1 and ET-RA aremore intense in vascular tissue and less intense in glomerularstructures. In contrast, subjects with IgAN show increased expressionsof ET-1 and ET-RA in the kidney. In that population, ET-1 expressionpositively correlates with proteinuria, which is at least partiallyameliorated by administration of ACE inhibitors. Indeed, the currentlytherapy for IgAN is optimization of antihypertensive and antiproteinuricagents (e.g., angiotensin converting enzyme inhibitors and/orangiotensin II receptor blockers), along with a course ofcorticosteroids, to inhibit disease progression. See, e.g., Penfold etal., Int. J. Nephrol. and Renovascular Dis. 11, pp. 137-148 (2017).However, these combinations of agents may exhibit significantdose-limiting side effects such as hyperkalemia, and furtherimmunosuppression may be necessary in more serious cases.

Clinically, IgAN is diagnosed by kidney biopsy indicating the presenceof mesangial cell proliferation and/or matrix expansion (or focalsegmental glomerular sclerosis in advanced stages) with predominantmesangial granular deposits of IgA (2+ or more) on immunofluorescence.This pathology is distinct from other progressive kidney diseases suchas diabetic nephropathy, which typically present with a diffusecapillary basement membrane thickening with peripheral hyalinePAS-positive nodules, with segmental or global glomerular sclerosis atadvanced stages, and thickened arterioles with hyaline deposits. See,e.g., Zanatta, et al., Renal Failure, 34(3), pp. 308-315 (2012).

Accordingly, in one aspect, provided herein is a method of treating IgAnephropathy, comprising administering a therapeutically effective amountof atrasentan, or a pharmaceutically acceptable salt thereof, to asubject in need thereof.

In some embodiments, the subject has not been previously diagnosed withone or more of diabetic nephropathy, HIV/AIDS, HIV-related nephropathy,prostate cancer, or acute kidney failure. In some embodiments, thesubject has not been previously diagnosed with any of diabeticnephropathy, HIV/AIDS, HIV-related nephropathy, prostate cancer, oracute kidney failure. In some embodiments, the subject has not beenpreviously diagnosed with diabetic nephropathy. In some embodiments, thesubject has not been previously diagnosed with HIV/AIDS. In someembodiments, the subject has not been previously diagnosed withHIV-related nephropathy. In some embodiments, the subject has not beenpreviously diagnosed with prostate cancer. In some embodiments, thesubject has not been previously diagnosed with acute kidney failure. Insome embodiments, the subject has not been previously diagnosed with oneor more of diabetic nephropathy, HIV/AIDS, HIV-related nephropathy,cancer, or acute kidney failure. In some embodiments, the subject hasnot been previously diagnosed with diabetes (i.e., Type 1 or Type 2diabetes). In some embodiments, the subject has been previouslydiagnosed with diabetes (i.e., Type 1 or Type 2 diabetes). In someembodiments, the subject has been previously diagnosed with diabetes,and has not been previously diagnosed with diabetic nephropathy. In someembodiments, the subject has not been previously diagnosed with Type 2diabetes. In some embodiments, the subject has been previously diagnosedwith Type 2 diabetes. In some embodiments, the subject has beenpreviously diagnosed with Type 2 diabetes, and has not been previouslydiagnosed with diabetic nephropathy.

In some embodiments, the subject is not currently diagnosed with cancer.In some embodiments, the subject is not currently being treated forcancer. In some embodiments, the cancer is lung cancer or prostatecancer.

In some embodiments, the subject does not have one or more of diabeticnephropathy, HIV/AIDS, HIV-related nephropathy, prostate cancer, oracute kidney failure. In some embodiments, the subject does not have anyof diabetic nephropathy, HIV/AIDS, HIV-related nephropathy, prostatecancer, or acute kidney failure. In some embodiments, the subject doesnot have diabetic nephropathy. In some embodiments, the subject does nothave HIV/AIDS. In some embodiments, the subject does not haveHIV-related nephropathy. In some embodiments, the subject does not havecancer. In some embodiments, the cancer is prostate cancer. In someembodiments, the cancer is lung cancer. In some embodiments, the subjectdoes not have acute kidney failure. In some embodiments, the subjectdoes not have one or more of diabetic nephropathy, HIV/AIDS, HIV-relatednephropathy, cancer, or acute kidney failure. In some embodiments, thecancer is lung cancer or prostate cancer. In some embodiments, thesubject has diabetes (i.e., Type 1 or Type 2 diabetes). In someembodiments, the subject does not have diabetes (i.e., Type 1 or Type 2diabetes). In some embodiments, the subject has diabetes, and does nothave diabetic nephropathy. In some embodiments, the subject has Type 2diabetes. In some embodiments, the subject does not have Type 2diabetes. In some embodiments, the subject has Type 2 diabetes, and doesnot have diabetic nephropathy.

In some embodiments, the subject does not suffer from one or more ofdiabetic nephropathy, HIV/AIDS, HIV-related nephropathy, prostatecancer, or acute kidney failure. In some embodiments, the subject doesnot suffer from any of diabetic nephropathy, HIV/AIDS, HIV-relatednephropathy, prostate cancer, or acute kidney failure. In someembodiments, the subject does not suffer from diabetic nephropathy. Insome embodiments, the subject does not suffer from HIV/AIDS. In someembodiments, the subject does not suffer from HIV-related nephropathy.In some embodiments, the subject does not suffer from cancer. In someembodiments, the cancer is prostate cancer. In some embodiments, thecancer is lung cancer. In some embodiments, the subject does not sufferfrom acute kidney failure. In some embodiments, the subject does notsuffer from one or more of diabetic nephropathy, HIV/AIDS, HIV-relatednephropathy, cancer, or acute kidney failure. In some embodiments, thecancer is lung cancer or prostate cancer. In some embodiments, thesubject does not suffer from diabetes (i.e., Type 1 or Type 2 diabetes).In some embodiments, the subject does suffer from diabetes (i.e., Type 1or Type 2 diabetes). In some embodiments, the subject does suffer fromdiabetes, such as Type 1 diabetes or Type 2 diabetes, but does notsuffer from diabetic nephropathy. In some embodiments, the subject doesnot suffer from Type 2 diabetes. In some embodiments, the subject doessuffer from Type 2 diabetes. In some embodiments, the subject doessuffer from Type 2 diabetes, but does not suffer from diabeticnephropathy.

In some embodiments, the subject is not being treated for one or more ofdiabetic nephropathy, HIV/AIDS, HIV-related nephropathy, prostatecancer, or acute kidney failure. In some embodiments, the subject is notbeing treated for any of diabetic nephropathy, HIV/AIDS, HIV-relatednephropathy, prostate cancer, or acute kidney failure. In someembodiments, the subject is not being treated for diabetic nephropathy.In some embodiments, the subject is not being treated for HIV/AIDS. Insome embodiments, the subject is not being treated for HIV-relatednephropathy. In some embodiments, the subject is not being treated forprostate cancer. In some embodiments, the subject is not being treatedfor acute kidney failure. In some embodiments, the subject is not beingtreated for one or more of diabetic nephropathy, HIV/AIDS, HIV-relatednephropathy, cancer, or acute kidney failure. In some embodiments, thecancer is lung cancer or prostate cancer. In some embodiments, thesubject is not being treated for diabetes (i.e., Type 1 or Type 2diabetes). In some embodiments, the subject is being treated fordiabetes (i.e., Type 1 or Type 2 diabetes). In some embodiments, thesubject is being treated for diabetes, such as Type 1 diabetes or Type 2diabetes, but is not being treated for diabetic nephropathy. In someembodiments, the subject is not being treated for Type 2 diabetes. Insome embodiments, the subject is being treated for Type 2 diabetes. Insome embodiments, the subject is being treated for Type 2 diabetes, butis not being treated for diabetic nephropathy.

In certain embodiments, the subject has been determined to havecontrolled serum glucose levels. In some embodiments, the subject withcontrolled serum glucose levels is not being treated for diabetes. Insome embodiments, the subject with controlled serum glucose levels isbeing treated for diabetes. In some embodiments, the subject withcontrolled serum glucose levels is not being treated for Type 2diabetes. In some embodiments, the subject with controlled serum glucoselevels is being treated for Type 2 diabetes. In some embodiments, thesubject has been determined to have controlled serum glucose levels;where the subject has not been diagnosed with one or more of HIV-relatednephropathy or acute kidney failure. For example, the subject has beendetermined to have a fasting serum glucose level of below about 130mg/dL, about 125 mg/dL, about 120 mg/dL, about 115 mg/dL, about 110mg/dL, about 105 mg/dL, about 100 mg/dL, about 95 mg/dL, about 90 mg/dL,about 85 mg/dL, about 80 mg/dL, or about 75 mg/dL, or any value inbetween. In certain embodiments, the subject has not been diagnosed withone or more of HIV-related nephropathy or acute kidney failure. Incertain embodiments, the subject has been determined to have controlledserum glucose levels as described anywhere herein; and the subject hasnot been diagnosed with one or more of HIV-related nephropathy or acutekidney failure. In certain embodiments, the subject has been determinedto have controlled serum glucose levels; and the subject has not beendiagnosed with one or more of HIV-related nephropathy or acute kidneyfailure.

In another aspect, provided herein is a method of decreasing renalinflammation and/or fibrosis in a subject having IgA nephropathy,comprising administering a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof, to a subjectin need thereof.

In some embodiments, the renal inflammation in the subject having IgAnephropathy is decreased by at least about 10% after treatment withatrasentan or a pharmaceutically acceptable salt thereof (e.g., aftertreatment for about 1 week, about 2 weeks, about 3 weeks, about 4 weeks,about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9weeks, about 10 weeks, about 20 weeks, about 30 weeks, about 40 weeks,about 50 weeks, about 60 weeks, about 70 weeks, about 80 weeks, about 90weeks, about 100 weeks, about 110 weeks, about 120 weeks, about 130weeks, about 140 weeks, about 150 weeks, about 160 weeks, about 170weeks, about 180 weeks, about 190 weeks, or about 200 weeks, or anyvalue in between). In certain embodiments, the renal inflammation in thesubject is decreased by at least about 20%, about 30%, about 40%, about50%, about 60%, about 70%, about 80%, about 90%, or about 95%, or anyvalue in between. In certain of the foregoing embodiments, the subjecthas been treated with atrasentan or a pharmaceutically acceptable saltthereof for between about 15 days and about 30 days.

In some embodiments, renal fibrosis in the subject having IgAnephropathy is decreased by at least about 10% after treatment withatrasentan or a pharmaceutically acceptable salt thereof (e.g., aftertreatment for about 1 week, about 2 weeks, about 3 weeks, about 4 weeks,about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9weeks, about 10 weeks, about 20 weeks, about 30 weeks, about 40 weeks,about 50 weeks, about 60 weeks, about 70 weeks, about 80 weeks, about 90weeks, about 100 weeks, about 110 weeks, about 120 weeks, about 130weeks, about 140 weeks, about 150 weeks, about 160 weeks, about 170weeks, about 180 weeks, about 190 weeks, or about 200 weeks, or anyvalue in between). In certain embodiments, the renal fibrosis in thesubject is decreased by at least about 20%, about 30%, about 40%, about50%, about 60%, about 70%, about 80%, about 90%, or about 95%, or anyvalue in between. In certain of the foregoing embodiments, the subjecthas been treated with atrasentan or a pharmaceutically acceptable saltthereof for between about 15 days and about 30 days.

In some embodiments, renal fibrosis in the subject having IgAnephropathy is decreased to less than about 50% of the cortical area ofthe affected kidney(s) after treatment with atrasentan or apharmaceutically acceptable salt thereof (e.g., after treatment forabout 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about10 weeks, about 20 weeks, about 30 weeks, about 40 weeks, about 50weeks, about 60 weeks, about 70 weeks, about 80 weeks, about 90 weeks,about 100 weeks, about 110 weeks, about 120 weeks, about 130 weeks,about 140 weeks, about 150 weeks, about 160 weeks, about 170 weeks,about 180 weeks, about 190 weeks, or about 200 weeks, or any value inbetween). In certain embodiments, about renal fibrosis in the subject isdecreased to less than about 40% of the cortical area. For example, insome embodiments, renal fibrosis in the subject is decreased to lessthan about 35%, about 30%, about 25%, about 20%, about 15%, or about10%, or any value in between, of the cortical area. In certain of theforegoing embodiments, the subject has been treated with atrasentan or apharmaceutically acceptable salt thereof for between about 15 days andabout 30 days.

In another aspect, provided herein is a method of decreasing theoccurrence of hematuria in a subject having IgA nephropathy, comprisingadministering a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof, to a subject in need thereof.

In some embodiments, the number of urinary red blood cells per highpowered (microscope) field (rbc/hpf) in the subject having IgAnephropathy is decreased by at least about 10% after treatment withatrasentan or a pharmaceutically acceptable salt thereof (e.g., aftertreatment for about 1 week, about 2 weeks, about 3 weeks, about 4 weeks,about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9weeks, about 10 weeks, about 20 weeks, about 30 weeks, about 40 weeks,about 50 weeks, about 60 weeks, about 70 weeks, about 80 weeks, about 90weeks, about 100 weeks, about 110 weeks, about 120 weeks, about 130weeks, about 140 weeks, about 150 weeks, about 160 weeks, about 170weeks, about 180 weeks, about 190 weeks, or about 200 weeks, or anyvalue in between). In certain embodiments, the urinary rbc/hpf in thesubject is decreased by at least about 20%, about 30%, about 40%, about50%, about 60%, about 70%, about 80%, about 90%, or about 95%, or anyvalue in between. In certain of the foregoing embodiments, the subjecthas been treated with atrasentan or a pharmaceutically acceptable saltthereof for between about 15 days and about 30 days.

In another aspect, provided herein is a method of stabilizing eGFR, in asubject having IgA nephropathy comprising administering atherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, to a subject in need thereof.

In some embodiments, provided herein is a method for reducing the rateof decrease of eGFR in a subject having IgA nephropathy, the methodcomprising administering a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof, to a subjectin need thereof. In some embodiments, the rate of decrease of eGFR ofthe subject is reduced by at least about 10% after treatment withatrasentan or a pharmaceutically acceptable salt thereof (e.g., aftertreatment for 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks,about 10 weeks, about 20 weeks, about 30 weeks, about 40 weeks, about 50weeks, about 60 weeks, about 70 weeks, about 80 weeks, about 90 weeks,about 100 weeks, about 110 weeks, about 120 weeks, about 130 weeks,about 140 weeks, about 150 weeks, about 160 weeks, about 170 weeks,about 180 weeks, about 190 weeks, or about 200 weeks, or any value inbetween). In some embodiments, the rate of decrease of eGFR of thesubject is reduced by at least about 20%; by at least about 30%; by atleast about 40%; by at least about 50%; by at least about 60%; by atleast about 70%; by at least about 80%; by at least about 90%; or by atleast about 95%; or any value in between. In certain of the foregoingembodiments, the subject has been treated with atrasentan or apharmaceutically acceptable salt thereof for between about 15 days andabout 30 days. In certain of the foregoing embodiments, the subject hasbeen treated with atrasentan or a pharmaceutically acceptable saltthereof for between about 6 months and about 1 year.

In some embodiments, the rate of decrease of eGFR of the subject havingIgA nephropathy is reduced to below about 10 mL/min/1.73 m² aftertreatment with atrasentan or a pharmaceutically acceptable salt thereof.For example, after treatment for 1 week, about 2 weeks, about 3 weeks,about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8weeks, about 9 weeks, about 10 weeks, about 20 weeks, about 30 weeks,about 40 weeks, about 50 weeks, about 60 weeks, about 70 weeks, about 80weeks, about 90 weeks, about 100 weeks, about 110 weeks, about 120weeks, about 130 weeks, about 140 weeks, about 150 weeks, about 160weeks, about 170 weeks, about 180 weeks, about 190 weeks, or about 200weeks, or any value in between. In certain embodiments, the rate ofdecrease of eGFR of the subject is reduced to below about 9 mL/min/1.73m², about 8 mL/min/1.73 m², about 7 mL/min/1.73 m², about 6 mL/min/1.73m², about 5 mL/min/1.73 m², about 4 mL/min/1.73 m², about 3 mL/min/1.73m², about 2 mL/min/1.73 m², about 1 mL/min/1.73 m², or about 0.75mL/min/1.73 m², or any value in between after treatment with atrasentan,or a pharmaceutically acceptable salt thereof, for between about 6months to about 1 year. The typical decline in eGFR with age, forexample, in a subject between about 20 to about 30 years of age, isabout 1 mL/min/1.73 m² per year.

In another aspect, provided herein is method of decreasing the number ofIgA-nephropathy associated disease flares in a subject having IgAnephropathy, comprising administering a therapeutically effective amountof atrasentan, or a pharmaceutically acceptable salt thereof, to asubject in need thereof. In some embodiments, the method decreasesdisease flares associated with hematuria. In some embodiments, themethod decreases disease flares associated with proteinuria. In someembodiments, the method decreases IgA-nephropathy associated diseaseflares which are associated with systemic manifestations. In someembodiments, the method decreases declines in eGFR as described anywhereherein. In some embodiments, the method decreases one or more of edema,fatigue, hematuria, or gross hematuria. In some embodiments, the methodpositively impacts disease progression.

In another aspect, provided herein is a method of delaying the onset ofESRD in a subject having IgA nephropathy, comprising administering atherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, to a subject in need thereof.

In some embodiments, the method increases the time between the diagnosisof IgA nephropathy in the subject and the time when eGFR of the subjectfalls below 15 mL/min/1.73 m². In certain embodiments, the methodincreases the time between the diagnosis of IgA nephropathy in thesubject and the time when eGFR of the subject falls below 15 mL/min/1.73m² by at least about 10%. For example, in some embodiments, the methodincreases the time between the diagnosis of IgA nephropathy in thesubject and the time when eGFR of the subject falls below 15 mL/min/1.73m² by at least about 20%, about 30%, about 40%, about 50%, about 60%,about 70%, about 80%, about 90%, about 100%, about 150%, about 200%,about 250%, about 300%, about 350%, about 400%, about 450%, or about500%, or any value in between.

In certain embodiments, the method increases the time between thediagnosis of IgA nephropathy in the subject and the time when eGFR ofthe subject falls below 15 mL/min/1.73 m² by at least about 1 year. Forexample, the method can delay the time when eGFR of the subject fallsbelow 15 mL/min/1.73 m² by at least about 1.5 years, 2 years, 2.5 years,3 years, 3.5 years, 4 years, 4.5 years, 5 years, 5.5 years, 6 years, 6.5years, 7 years, 7.5 years, 8 years, 8.5 years, 9 years, 9.5 years, 10years, 11 years, 12 years, 13 years, 15 years, 15 years, 16 years, 17years, 18 years, 19 years, or 20 years.

In another aspect, provided herein is a method of decreasing proteinuriain a subject having IgA nephropathy, comprising administering atherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, to a subject in need thereof.

In some embodiments, the amount of proteins (e.g., albumin) in the urineof the subject having IgA nephropathy is reduced by at least about 10%after treatment with atrasentan or a pharmaceutically acceptable saltthereof (e.g., after treatment for 1 week, about 2 weeks, about 3 weeks,about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8weeks, about 9 weeks, about 10 weeks, about 20 weeks, about 30 weeks,about 40 weeks, about 50 weeks, about 60 weeks, about 70 weeks, about 80weeks, about 90 weeks, about 100 weeks, about 110 weeks, about 120weeks, about 130 weeks, about 140 weeks, about 150 weeks, about 160weeks, about 170 weeks, about 180 weeks, about 190 weeks, or about 200weeks, or any value in between). In some embodiments, the amount ofproteins in the urine of the subject is reduced by at least about 15%,about 20%, about 25%, about 30%, about 35%, about 40%, about 50%, about55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%,about 90%, or about 95%, or any value in between. In certain of theforegoing embodiments, the subject has been treated with atrasentan or apharmaceutically acceptable salt thereof for between about 15 days andabout 30 days.

In certain embodiments, the amount of proteins (e.g., albumin) in theurine of the subject having IgA nephropathy is reduced by about 20% toabout 80% after between about 2 days and about 30 days of treatment withatrasentan, or a pharmaceutically acceptable salt thereof. In certainembodiments, the amount of proteins (e.g., albumin) in the urine of thesubject having IgA nephropathy is reduced by about 20% to about 80%after between about 15 days and about 30 days of treatment withatrasentan, or a pharmaceutically acceptable salt thereof. In certain ofthese embodiments, the amount of proteins in the urine of the subject isreduced by about 25% to about 80%. In certain of these embodiments, theamount of proteins in the urine of the subject is reduced by about 30%to about 80%. In certain of these embodiments, the amount of proteins inthe urine of the subject is reduced by about 35% to about 80%. Incertain of these embodiments, the amount of proteins in the urine of thesubject is reduced by about 40% to about 80%. In certain of theseembodiments, the amount of proteins in the urine of the subject isreduced by about 45% to about 80%. In certain of these embodiments, theamount of proteins in the urine of the subject is reduced by about 50%to about 80%. In the aforementioned embodiments, the reduction of theamount of proteins (e.g., albumin) in the urine of the subject havingIgA nephropathy is relative to the amount of proteins (e.g., albumin) inthe urine prior to initiation of treatment with atrasentan, or apharmaceutically acceptable salt thereof.

In some embodiments, the amount of proteins (e.g., albumin) in the urineof the subject having IgA nephropathy is reduced by about 100 mg/dL toabout 3,000 mg/dL after treatment with atrasentan or a pharmaceuticallyacceptable salt thereof (e.g., after treatment for 1 week, about 2weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 20 weeks,about 30 weeks, about 40 weeks, about 50 weeks, about 60 weeks, about 70weeks, about 80 weeks, about 90 weeks, about 100 weeks, about 110 weeks,about 120 weeks, about 130 weeks, about 140 weeks, about 150 weeks,about 160 weeks, about 170 weeks, about 180 weeks, about 190 weeks, orabout 200 weeks, or any value in between). In certain embodiments, theamount of proteins in the urine of the subject is reduced by about 100mg/dL to about 2,500 mg/dL. In certain embodiments, the amount ofproteins in the urine of the subject is reduced by about 100 mg/dL toabout 2,000 mg/dL. In certain embodiments, the amount of proteins in theurine of the subject is reduced by about 100 mg/dL to about 1,500 mg/dL.In certain embodiments, the amount of proteins in the urine of thesubject is reduced by about 100 mg/dL to about 1,000 mg/dL. In certainembodiments, the amount of proteins in the urine of the subject isreduced by about 100 mg/dL to about 500 mg/dL. In certain embodiments,the amount of proteins in the urine of the subject is reduced by about100 mg/dL to about 400 mg/dL. In certain embodiments, the amount ofproteins in the urine of the subject is reduced by about 100 mg/dL toabout 300 mg/dL. In certain embodiments, the amount of proteins in theurine of the subject is reduced by about 100 mg/dL to about 200 mg/dL.In certain embodiments, the amount of proteins in the urine of thesubject is reduced by about 500 mg/dL to about 2,500 mg/dL. In certainembodiments, the amount of proteins in the urine of the subject isreduced by about 500 mg/dL to about 2,000 mg/dL. In certain embodiments,the amount of proteins in the urine of the subject is reduced by about500 mg/dL to about 1,500 mg/dL. In certain embodiments, the amount ofproteins in the urine of the subject is reduced by about 500 mg/dL toabout 1,000 mg/dL. In certain embodiments, the amount of proteins in theurine of the subject is reduced by about 500 mg/dL to about 900 mg/dL.In certain embodiments, the amount of proteins in the urine of thesubject is reduced by about 500 mg/dL to about 800 mg/dL. In certainembodiments, the amount of proteins in the urine of the subject isreduced by about 600 mg/dL to about 900 mg/dL. In certain embodiments,the amount of proteins in the urine of the subject is reduced by about700 mg/dL to about 900 mg/dL. In certain embodiments, the amount ofproteins in the urine of the subject is reduced by about 1,000 mg/dL toabout 2,000 mg/dL. In certain of the foregoing embodiments, the subjecthas been treated with atrasentan or a pharmaceutically acceptable saltthereof for between about 15 days and about 30 days. In theaforementioned embodiments, the reduction of the amount of proteins(e.g., albumin) in the urine of the subject having IgA nephropathy isrelative to the amount of proteins (e.g., albumin) in the urine prior toinitiation of treatment with atrasentan, or a pharmaceuticallyacceptable salt thereof.

In certain embodiments, the amount of proteins (e.g., albumin) in theurine of the subject having IgA nephropathy is reduced by about 100mg/dL to about 500 mg/dL after between about 15 days and about 30 daysof treatment with atrasentan, or a pharmaceutically acceptable saltthereof. In certain embodiments, the amount of proteins in the urine ofthe subject is reduced by about 200 mg/dL to about 500 mg/dL afterbetween about 15 days and about 30 days of treatment with atrasentan, ora pharmaceutically acceptable salt thereof. In certain embodiments, theamount of proteins in the urine of the subject is reduced by about 300mg/dL to about 500 mg/dL after between about 15 days and about 30 daysof treatment with atrasentan, or a pharmaceutically acceptable saltthereof. In the aforementioned embodiments, the reduction of the amountof proteins (e.g., albumin) in the urine of the subject having IgAnephropathy is relative to the amount of proteins (e.g., albumin) in theurine prior to initiation of treatment with atrasentan, or apharmaceutically acceptable salt thereof.

In certain embodiments, the amount of proteins (e.g., albumin) in theurine of the subject having IgA nephropathy is reduced by about 500mg/dL to about 900 mg/dL after between about 15 days and about 30 daysof treatment with atrasentan, or a pharmaceutically acceptable saltthereof. In certain embodiments, the amount of proteins in the urine ofthe subject is reduced by about 600 mg/dL to about 900 mg/dL afterbetween about 15 days and about 30 days of treatment with atrasentan, ora pharmaceutically acceptable salt thereof. In certain embodiments, theamount of proteins in the urine of the subject is reduced by about 700mg/dL to about 900 mg/dL after between about 15 days and about 30 daysof treatment with atrasentan, or a pharmaceutically acceptable saltthereof. In the aforementioned embodiments, the reduction of the amountof proteins (e.g., albumin) in the urine of the subject having IgAnephropathy is relative to the amount of proteins (e.g., albumin) in theurine prior to initiation of treatment with atrasentan, or apharmaceutically acceptable salt thereof.

In some embodiments, the subject having IgA nephropathy has a reducedlevel of proteins (e.g., albumin) in the urine of below about 1.0gram/day after treatment with atrasentan or a pharmaceuticallyacceptable salt thereof (e.g., after treatment for 1 week, 2 weeks, 3weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks,20 weeks, 30 weeks, 40 weeks, 50 weeks, 60 weeks, 70 weeks, 80 weeks, 90weeks, 100 weeks, 110 weeks, 120 weeks, 130 weeks, 140 weeks, 150 weeks,160 weeks, 170 weeks, 180 weeks, 190 weeks, or 200 weeks). In certainembodiments, the subject has a reduced level of proteins in the urine ofbelow about 0.9 gram/day. In certain embodiments, the subject has areduced level of proteins in the urine of below about 0.8 gram/day. Incertain embodiments, the subject has a reduced level of proteins in theurine of below about 0.7 gram/day. In certain embodiments, the subjecthas a reduced level of proteins in the urine of below about 0.6gram/day. In certain embodiments, the subject has a reduced level ofproteins in the urine of below about 0.5 gram/day. In certainembodiments, the subject has a reduced level of proteins in the urine ofbelow about 0.4 gram/day. In certain embodiments, the subject has areduced level of proteins in the urine of below about 0.3 gram/day. Incertain embodiments, the subject has a reduced level of proteins in theurine of below about 0.2 gram/day. In certain of the foregoingembodiments, the subject has been treated with atrasentan or apharmaceutically acceptable salt thereof for between about 15 days andabout 30 days. In the aforementioned embodiments, the reduction of theamount of proteins (e.g., albumin) in the urine of the subject havingIgA nephropathy is relative to the amount of proteins (e.g., albumin) inthe urine prior to initiation of treatment with atrasentan, or apharmaceutically acceptable salt thereof.

In another aspect, provided herein is a method of decreasing fatigue ina subject having IgA nephropathy, comprising administering atherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, to a subject in need thereof. In someembodiments, the subject has been determined not to suffer from one ormore of diabetic nephropathy, HIV-related nephropathy, prostate cancer,or acute kidney failure. In certain embodiments, the subject has beendetermined not to suffer from diabetic nephropathy. In certainembodiments, the subject has been determined not to suffer fromHIV-related neuropathy. In certain embodiments, the subject has beendetermined not to suffer from prostate cancer. In certain embodiments,the subject has been determined not to suffer from acute kidney failure.

In some embodiments, the fatigue of the subject having IgA nephropathyis reduced by about 5% to about 80% after treatment with atrasentan or apharmaceutically acceptable salt thereof (e.g., after treatment for 1week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks,about 20 weeks, about 30 weeks, about 40 weeks, about 50 weeks, about 60weeks, about 70 weeks, about 80 weeks, about 90 weeks, about 100 weeks,about 110 weeks, about 120 weeks, about 130 weeks, about 140 weeks,about 150 weeks, about 160 weeks, about 170 weeks, about 180 weeks,about 190 weeks, or about 200 weeks, or any value in between). Incertain embodiments, the fatigue is reduced by about 10% to about 75%.In certain embodiments, the fatigue is reduced by about 10% to about70%. In certain embodiments, the fatigue is reduced by about 10% toabout 65%. In certain embodiments, the fatigue is reduced by about 10%to about 60%. In certain embodiments, the fatigue is reduced by about10% to about 55%. In certain embodiments, the fatigue is reduced byabout 10% to about 50%. In certain embodiments, the fatigue is reducedby about 10% to about 45%. In certain embodiments, the fatigue isreduced by about 10% to about 40%. In certain embodiments, the fatigueis reduced by about 10% to about 35%. In certain embodiments, thefatigue is reduced by about 10% to about 30%. In certain embodiments,the fatigue is reduced by about 10% to about 25%. In certainembodiments, the fatigue is reduced by about 10% to about 20%. Incertain embodiments, the fatigue is reduced by about 10% to about 15%.In certain embodiments, the fatigue is reduced by about 20% to about75%. In certain embodiments, the fatigue is reduced by about 20% toabout 70%. In certain embodiments, the fatigue is reduced by about 20%to about 65%. In certain embodiments, the fatigue is reduced by about20% to about 60%. In certain embodiments, the fatigue is reduced byabout 20% to about 55%. In certain embodiments, the fatigue is reducedby about 20% to about 50%. In certain embodiments, the fatigue isreduced by about 20% to about 45%. In certain embodiments, the fatigueis reduced by about 20% to about 40%. In certain embodiments, thefatigue is reduced by about 20% to about 35%. In certain embodiments,the fatigue is reduced by about 20% to about 30%. In certainembodiments, the fatigue is reduced by about 30% to about 75%. Incertain embodiments, the fatigue is reduced by about 30% to about 70%.In certain embodiments, the fatigue is reduced by about 30% to about65%. In certain embodiments, the fatigue is reduced by about 30% toabout 60%. In certain embodiments, the fatigue is reduced by about 30%to about 55%. In certain embodiments, the fatigue is reduced by about30% to about 50%. In certain embodiments, the fatigue is reduced byabout 30% to about 45%. In certain embodiments, the fatigue is reducedby about 30% to about 40%. In certain embodiments, the fatigue isreduced by about 40% to about 75%. In certain embodiments, the fatigueis reduced by about 40% to about 70%. In certain embodiments, thefatigue is reduced by about 40% to about 65%. In certain embodiments,the fatigue is reduced by about 40% to about 60%. In certainembodiments, the fatigue is reduced by about 40% to about 55%. Incertain embodiments, the fatigue is reduced by about 40% to about 50%.In certain embodiments, the fatigue is reduced by about 50% to about75%. In certain embodiments, the fatigue is reduced by about 50% toabout 70%. In certain embodiments, the fatigue is reduced by about 50%to about 65%. In certain embodiments, the fatigue is reduced by about50% to about 60%. In certain of the foregoing embodiments, the subjecthas been treated with atrasentan or a pharmaceutically acceptable saltthereof for between about 15 days and about 30 days. In certainembodiments, the decrease in fatigue comprises a decrease in the scoreon one or more of the Fatigue Severity Scale, the Chalder Fatigue Scale,the FACIT Fatigue Scale, the Brief Fatigue Inventory, the FACT-FSubscale, Global Vigor and Affect, the May and Kline AdjectiveChecklist, the Pearson-Byars Fatigue Feeling Checklist, the RhotenFatigue Scale, the Schedule of Fatigue and Anergia, the Visual AnalogScale, or the Checklist Individual Strength. In the aforementionedembodiments, the reduction of fatigue experienced by the subject havingIgA nephropathy is relative to the fatigue experienced by the subjectprior to initiation of treatment with atrasentan, or a pharmaceuticallyacceptable salt thereof. In some embodiments, the decrease in fatiguecomprises a decrease in the score on the Brief Fatigue Inventory.

Subject Selection

The subject having IgA nephropathy as described anywhere herein can bediagnosed using one or more methods known in the art. Non-limitingexamples include: kidney biopsy, detecting galactose-deficient IgA(e.g., Gd-IgA1), detecting anti-glycan antibodies, detecting depositionof IgA-immune complexes in the kidney, or a combination of any of theforegoing. In some embodiments, the diagnosis of IgA nephropathycomprises detecting deposition of IgA-immune complexes in the kidney. Incertain embodiments, the diagnosis of IgA nephropathy comprises a kidneybiopsy. In certain embodiments, the diagnosis of IgA nephropathycomprises detecting galactose-deficient IgA. In certain embodiments, thediagnosis of IgA nephropathy comprises detecting anti-glycan antibodies(e.g., KM55). In certain embodiments, the diagnosis of IgA nephropathycomprises a kidney biopsy followed by detecting deposition of IgA-immunecomplexes in the kidney (for example, by light microscopy and/orimmunofluorescence microscopy).

In some embodiments, the presence and/or level of a particular proteinin a subject is determined prior to administration of atrasentan, or apharmaceutically acceptable salt thereof. For example, serum levels ofGd-IgA1, serum levels of autoantibodies specific for Gd-IgA1, and/orserum and/or urine levels of IgA1-containing immune complexes. See,e.g., Knoppova, et al., Front. Immunol., Vol. 17, Art. 117 (2016), whichis hereby incorporated by reference in its entirety. In someembodiments, the subject has Gd-IgA levels in the 90^(th) percentile orabove prior to administration of atrasentan or a pharmaceuticallyacceptable salt thereof. In some embodiments, the subject has Gd-IgAlevels in the 95^(th) percentile or above prior to administration ofatrasentan or a pharmaceutically acceptable salt thereof. In someembodiments, the subject's Gd-IgA levels decrease to below the 90^(th)percentile after treatment with atrasentan, or a pharmaceuticallyacceptable salt thereof, for between about 6 months to 1 year.

In certain embodiments, the subject has mesangial cellularity in about≥50% (e.g., about ≥60%, about ≥70%, or about ≥80%) of the glomeruli,wherein mesangial cellularity is defined as more than four mesangialcells in any mesangial area of a glomerulus. In certain embodiments,endocapillary hypercellularity is present in the subject, whereinendocapillary hypercellularity is defined as hypercellularity due to anincreased number of cells within glomerular capillary lumina. In certainembodiments, segmental sclerosis is present in the subject, whereinsegmental sclerosis is defined as adhesion or sclerosis (obliteration ofcapillary lumina by matrix) in part of but not the whole glomerulartuft. In certain embodiments, the subject has tubularatrophy/interstitial fibrosis in about ≥50% (e.g., about ≥60%, about≥65%, about ≥70%, about ≥75%, or about ≥80%) of the cortical area,wherein tubular atrophy/interstitial fibrosis is defined as theestimated percentage of cortical area showing tubular atrophy orinterstitial fibrosis. In certain embodiments, the subject has crescentspresent on the glomeruli. In certain of these embodiments, the subjecthas crescents present on below about 25% (e.g., below about 20%, about15%, about 10%, or about 5%) of the glomeruli. In certain embodiments,the subject has a MEST-C score of M1; E1; S1; T1 or T2; and/or C0 or C1under the Oxford MEST-C classification system. The Oxford MEST-Cclassification system is defined in Kidney International (2009) 76,546-556 and Nature Reviews Nephrology (2017) 13, 385-386, each of whichis incorporated herein by reference in its entirety (Also see: KidneyResearch and Clinical Practice (2016) 35, 197-203; and IgA Nephropathyin Medscape (accessed Nov. 4, 2019), each of which is incorporatedherein by reference in its entirety)).

In some embodiments, the subject is at a high risk of progression toESRD. In certain of these embodiments, the subject is excreting anaverage of about 1 gram or more of protein in the urine per day for atleast about 3 months prior to the first administration of atrasentan, ora pharmaceutically acceptable salt thereof. In certain embodiments, thesubject has an average eGFR ≤60 mL/min/1.73 m² (e.g., about ≤55, about≤50, about ≤45, about ≤40, about ≤35) for at least about 3 months priorto the first administration of atrasentan, or a pharmaceuticallyacceptable salt thereof. In certain of these embodiments, the subjecthas eGFR >30 mL/min/1.73 m² prior to the first administration ofatrasentan or a pharmaceutically acceptable salt thereof.

In some embodiments, the subject is excreting an average of about 1 gramor more of protein in the urine per day for at least about 3 months(e.g., at least about 4 months, at least about 5 months, at least about6 months, at least about 7 months, at least about 8 months, at leastabout 9 months, at least about 10 months, at least about 11 months, atleast about a year, at least about 1.5 years, or at least about 2 years)prior to the first administration of atrasentan, or a pharmaceuticallyacceptable salt thereof. For example, the subject can be excreting anaverage of about 1.1 gram, 1.2 grams, 1.3 grams, 1.4 grams, 1.5 grams,1.6 grams, 1.7 grams, 1.8 grams, 1.9 grams, 2.0 grams, 2.1 grams, 2.2grams, 2.3 grams, 2.4 grams, 2.5 grams, 2.6 grams, 2.7 grams. 2.8 grams,2.9 grams, 3.0 grams, 3.1 grams, 3.2 grams, 3.3 grams, 3.4 grams, 3.5grams, 5 grams, or 7.5 grams, or 10 grams, or any value in between, ofprotein in the urine per day for at least 3 months prior to the firstadministration of atrasentan, or a pharmaceutically acceptable saltthereof.

In some embodiments, the subject is excreting an average of from about0.3 grams to about 2 grams of protein in the urine per day for at leastabout 3 months (e.g., at least about 4 months, at least about 5 months,at least about 6 months, at least about 7 months, at least about 8months, at least about 9 months, at least about 10 months, at leastabout 11 months, at least about a year, at least about 1.5 years, or atleast about 2 years) prior to the first administration of atrasentan, ora pharmaceutically acceptable salt thereof. For example, the subject canbe excreting from about 0.3 grams to 0.5 grams, 0.5 grams to 1 gram,from about 0.5 grams to 1.5 grams, from about 1 gram to 1.5 grams, orfrom about 1.5 grams to 2 grams of protein in the urine per day for atleast 3 months.

In some embodiments, the subject is excreting at least about 1 gram ofprotein in the urine per day on at least two of three consecutivemeasurements a year prior to the first administration of atrasentan or apharmaceutically acceptable salt thereof. For example, the subject canbe excreting about 1.1 grams, 1.2 grams, 1.3 grams, 1.4 grams, 1.5grams, 1.6 grams, 1.7 grams, 1.8 grams, 1.9 grams, 2.0 grams, 2.1 grams,2.2 grams, 2.3 grams, 2.4 grams, 2.5 grams, 2.6 grams, 2.7 grams. 2.8grams, 2.9 grams, 3.0 grams, 3.1 grams, 3.2 grams, 3.3 grams, 3.4 grams,3.5 grams, 5 grams, or 7.5 grams, or 10 grams of protein, or any valuein between, in the urine per day on at least two of three consecutivemeasurements a year prior to the first administration of atrasentan or apharmaceutically acceptable salt thereof.

In some embodiments, the subject has an UACR value of at least about 300mg/g for at least three months prior to the first administration ofatrasentan or a pharmaceutically acceptable salt thereof, for example,300 mg/g to about 5,000 mg/g. In some embodiments, the subject has anUACR value of about 800 mg/g for at least three months prior to thefirst administration of atrasentan or a pharmaceutically acceptable saltthereof, for example, 800 mg/g to about 5,000 mg/g. In some embodiments,the subject has an UACR value of at least about 500 mg/g, about 600mg/g, about 700 mg/g, about 800 mg/g, about 900 mg/g, about 1,000 mg/g,about 1,500 mg/g, about 2,000 mg/g, about 2,500 mg/g, about 3,000 mg/g,about 3,500 mg/g, about 4,000 mg/g, about 4,500 mg/g, or about 5,000mg/g, or any value in between, for at least three months prior to thefirst administration of atrasentan or a pharmaceutically acceptable saltthereof.

In some embodiments, the subject has a decrease in UACR value of atleast about 30% relative to the subject's average UACR value for atleast three months prior to the first administration of atrasentan or apharmaceutically acceptable salt thereof, for example, a decrease ofabout 30% to about 100%, relative to the subject's average UACR valuefor at least three months prior to the first administration ofatrasentan or a pharmaceutically acceptable salt thereof. In someembodiments, the subject has a decrease in UACR value of at least about30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%,or about 100%, or any value in between, relative to the subject'saverage UACR value for at least three months prior to the firstadministration of atrasentan or a pharmaceutically acceptable saltthereof. In some embodiments, the subject having a decrease in UACRvalue does not also experience significant sodium retention and/orsignificant fluid retention. In some embodiments, significant fluidretention can be about 1 kg to about 4 kg over six weeks, for example,about 4 kg, about 3.5 kg, about 3 kg, about 2.5 kg, about 2 kg, about1.5 kg, or about 1 kg, or any value in between over 6 weeks. In someembodiments, a subject having significant fluid retention exhibitsclinical symptoms of edema.

In certain embodiments, the subject has an average eGFR of about 20 toabout 90 mL/min/1.73 m² for at least about 3 months prior to the firstadministration of atrasentan, or a pharmaceutically acceptable saltthereof (e.g., about 3 months, about 4 months, about 5 months, about 6months, about 7 months, about 8 months, about 9 months, about 10 months,about 11 months, about 12 months, about 1.5 years, or about 2 years).For example, about 20 to about 50 mL/min/1.73 m²; about 30 to about 60mL/min/1.73 m²; about 40 to about 70 mL/min/1.73 m²; about 50 to about80 mL/min/1.73 m²; or about 60 to about 90 mL/min/1.73 m²; for at leastabout 3 months prior to the first administration of atrasentan, or apharmaceutically acceptable salt thereof. In some embodiments, thesubject has an average eGFR ≤60 mL/min/1.73 m² for at least about 3months prior to the first administration of atrasentan, or apharmaceutically acceptable salt thereof. In certain embodiments, thesubject has an average eGFR ≤55 mL/min/1.73 m² for at least about 3months. In certain embodiments, the subject has an average eGFR ≤50mL/min/1.73 m² for at least about 3 months. In certain embodiments, thesubject has an average eGFR ≤45 mL/min/1.73 m² for at least about 3months. In certain embodiments, the subject has an average eGFR ≤40mL/min/1.73 m² for at least about 3 months. In certain embodiments, thesubject has an average eGFR ≤35 mL/min/1.73 m² for at least about 3months. In certain embodiments, the subject has an average eGFR ≤25mL/min/1.73 m² for at least about 3 months. In certain embodiments, thesubject has an average eGFR ≤20 mL/min/1.73 m² for at least about 3months. In certain of the foregoing embodiments, the subject has anaverage eGFR between about 30 mL/min/1.73 m² and about 60 mL/min/1.73 m²for at least 3 months before the administration of atrasentan or apharmaceutically acceptable salt thereof. For example, the subject canhave an average eGFR of between about 30 mL/min/1.73 m² and about 55mL/min/1.73 m², between about 30 mL/min/1.73 m² and about 50 mL/min/1.73m², between about 30 mL/min/1.73 m² and about 45 mL/min/1.73 m², orbetween about 30 mL/min/1.73 m² and about 40 mL/min/1.73 m².

In certain embodiments, the subject has an average eGFR of about 30mL/min/1.73 m² to about 45 mL/min/1.73 m², for example, about ≤45, about≤40, about ≤35 or about ≤30, for at least about 3 months (e.g., at leastabout 4 months, at least about 5 months, at least about 6 months, atleast about 7 months, at least about 8 months, at least about 9 months,at least about 10 months, at least about 11 months, at least about ayear, at least about 1.5 years, or at least about 2 years) prior to thefirst administration of atrasentan, or a pharmaceutically acceptablesalt thereof. In some embodiments, the subject has an average eGFR ofabout 25 mL/min/1.73 m² to about 75 mL/min/1.73 m² for at least about 3months prior to the first administration of atrasentan, or apharmaceutically acceptable salt thereof. For example, about 25mL/min/1.73 m², about 30 mL/min/1.73 m², about 35 mL/min/1.73 m², about40 mL/min/1.73 m², about 45 mL/min/1.73 m², about 50 mL/min/1.73 m²,about 55 mL/min/1.73 m², about 60 mL/min/1.73 m², about 65 mL/min/1.73m², about 70 mL/min/1.73 m², about 75 mL/min/1.73 m², or any value inbetween, for at least about 3 months prior to the first administrationof atrasentan, or a pharmaceutically acceptable salt thereof.

In some embodiments, the subject has an average HbA1c of about 4% toabout 6% for at least about 3 months (e.g., at least about 4 months, atleast about 5 months, at least about 6 months, at least about 7 months,at least about 8 months, at least about 9 months, at least about 10months, at least about 11 months, at least about a year, at least about1.5 years, or at least about 2 years) prior to the first administrationof atrasentan, or a pharmaceutically acceptable salt thereof. Forexample, the subject can have an average HbA1c of about 4.2%, about4.4%, about 4.6%, about 4.8%, about 5.0%, about 5.2%, about 5.4%, about5.6%, about 5.8%, or about 6%, or any value in between.

In some embodiments, the subject has an average fasting blood glucoselevel of about 125 mg/dL or less for at least about 3 months (e.g., atleast about 4 months, at least about 5 months, at least about 6 months,at least about 7 months, at least about 8 months, at least about 9months, at least about 10 months, at least about 11 months, at leastabout a year, at least about 1.5 years, or at least about 2 years) priorto the first administration of atrasentan, or a pharmaceuticallyacceptable salt thereof. For example, the subject can have an averagefasting blood glucose level of about 120 mg/dL, about 115 mg/dL, about110 mg/dL, about 105 mg/dL, about 100 mg/dL, about 95 mg/dL, about 90mg/dL, about 85 mg/dL, about 80 mg/dL, or about 75 mg/dL, or any valuein between.

In some embodiments, the subject maintains a potassium level within thenormal physiologic range. In certain embodiments, the subject maintainsa potassium level within the normal physiologic range for at least about3 months (e.g., at least about 4 months, at least about 5 months, atleast about 6 months, at least about 7 months, at least about 8 months,at least about 9 months, at least about 10 months, at least about 11months, at least about a year, at least about 1.5 years, or at leastabout 2 years) prior to the first administration of atrasentan or apharmaceutically acceptable salt thereof. In certain embodiments, thesubject maintains a potassium level within 3.5 to 5.2 mEq/L. Forexample, the subject maintains an average potassium level at about 3.5mEq/L, about 3.6 mEq/L, about 3.7 mEq/L, about 3.8 mEq/L, about 3.9,about mEq/L, about 4.0 mEq/L, about 4.1 mEq/L, about 4.2 mEq/L, about4.3 mEq/L, about 4.4 mEq/L, about 4.5 mEq/L, about 4.6 mEq/L, about 4.7mEq/L, about 4.8 mEq/L, about 4.9 mEq/L, about 5.0 mEq/L, about 5.1mEq/L, or about 5.2 mEq/L, or any value in between.

In some embodiments, the subject maintains a sodium level within thenormal physiologic range. In certain embodiments, the subject maintainsa potassium level within the normal physiologic range for at least about3 months (e.g., at least about 4 months, at least about 5 months, atleast about 6 months, at least about 7 months, at least about 8 months,at least about 9 months, at least about 10 months, at least about 11months, at least about a year, at least about 1.5 years, or at leastabout 2 years) prior to the first administration of atrasentan or apharmaceutically acceptable salt thereof. In certain embodiments, thesubject maintains a sodium level within 135 to 145 mEq/L. For example,the subject maintains an average sodium level of about 135 mEq/L, about136 mEq/L, about 137 mEq/L, about 138 mEq/L, about 139 mEq/L, about 140mEq/L, about 141 mEq/L, about 142 mEq/L, about 143 mEq/L, about 144mEq/L, about or 145 mEq/L, or any value in between.

In some embodiments, the subject has ALT/AST levels during theadministration of atrasentan, or a pharmaceutically acceptable saltthereof, that are about the same as the ALT/AST levels prior to thefirst administration of atrasentan, or a pharmaceutically acceptablesalt thereof. For example, the subject has ALT/AST levels during theadministration of atrasentan or a pharmaceutically acceptable saltthereof within about 25%, about 20%, about 15%, about 10%, about 5%,about or 2.5%, or any value in between, of the levels prior to the firstadministration of atrasentan, or a pharmaceutically acceptable saltthereof.

In some embodiments, the subject has bilirubin levels during theadministration of atrasentan, or a pharmaceutically acceptable saltthereof, that are about the same as the bilirubin levels prior to thefirst administration of atrasentan, or a pharmaceutically acceptablesalt thereof. For example, the subject has bilirubin levels during theadministration of atrasentan or a pharmaceutically acceptable saltthereof within about 25%, about 20%, about 15%, about 10%, about 5%, orabout 2.5%, or any value in between, of the levels prior to the firstadministration of atrasentan, or a pharmaceutically acceptable saltthereof.

In some embodiments, the fluid retention in the subject is manageablewith diuretics (e.g., during the treatment with atrasentan or apharmaceutically acceptable salt thereof and/or prior to the firstadministration of atrasentan or a pharmaceutically acceptable saltthereof). For example, the fluid retention can be less than about 3kilograms (kg) of weight gain over 6 weeks. In some embodiments, thefluid retention is less than about 4 kg, about 3.5 kg, about 3 kg, about2.5 kg, about 2 kg, about 1.5 kg, or about 1 kg, or any value in betweenover 6 weeks.

In some embodiments, the subject undergoes surgery, and/or otherregimens prior to, substantially at the same time as, or following theadministration of atrasentan, or a pharmaceutically acceptable saltthereof, as disclosed herein. In some embodiments, the subject isadministered other chemical and/or biological therapeutic agents priorto, substantially at the same time as, or following the administrationof atrasentan, or a pharmaceutically acceptable salt thereof, asdisclosed herein.

In some embodiments, the subject has been receiving one or moreinhibitors of the renin-angiotensin system for at least about 60 weeksprior to the first administration of atrasentan or a pharmaceuticallyacceptable salt thereof. For example, in some embodiments, the subjecthas been receiving one or more inhibitors of the renin-angiotensinsystem for at least about 12 weeks, about 24 weeks, about 48 weeks, orabout 60 weeks, or any value in between, prior to the firstadministration of atrasentan or a pharmaceutically acceptable saltthereof.

In some embodiments, the subject has been receiving a maximallytolerated stable dose of the one or more renin-angiotensin systeminhibitors. For example, the subject can be receiving a maximallytolerated stable dose of the one or more renin-angiotensin systeminhibitor for at least about 12 weeks, about 14 weeks, about 16 weeks,about 18 weeks, about 20 weeks, about 25 weeks, about 30 weeks, about 35weeks, about 40 weeks, about 45 weeks, or about 50 weeks, or any valuein between, prior to the first administration of atrasentan or apharmaceutically acceptable salt thereof. In some embodiments, the oneor more inhibitors of the renin-angiotensin system is selected from thegroup consisting of angiotensin converting enzyme (ACE) inhibitors,angiotensin II receptor blockers (ARBs), renin inhibitors, andaldosterone antagonists. For example, the one or more inhibitors of therenin-angiotensin system can be ACE inhibitor, ARB, or a combinationthereof, wherein the ACE inhibitor or ARB can be described anywhereherein. For example, the ACE inhibitor can be selected from: quinapril,fosinopril perindopril, captopril, enalapril, enalaprilat, ramipril,cilazapril, delapril, fosenopril, zofenopril, indolapril, benazepril,lisinopril, spirapril, trandolapril, perindep, pentopril, moexipril,rescinnamine, and pivopril. For example, the ARB can be selected from:candesartan, candesartan cilexetil, eprosartan, irbesartan, losartan,olmesartan, olmesartan medoxomil, telmisartan, valsartan, azilsartanmedoxomil, and BRA-657.

In some embodiments, the subject is also being administered one or moreadditional agents. In some embodiments, the one or more additionalagents are selected from calcineurin inhibitors, proteasome inhibitors,aminoquinolines, complement inhibitors, B-cell inhibitors, cytotoxicagents, mTOR inhibitors, and steroids. In some embodiments, the dosageof the one or more additional agents is decreased after between about 15days to about 30 days of treatment with atrasentan, or apharmaceutically acceptable salt thereof. In some embodiments, the oneor more additional agents are immunosuppressants.

In some embodiments, the subject is not currently receiving one or moreadditional agents. In certain embodiments, the subject has not used oneor more additional agent for two or more weeks within the 6 months priorto the first administration of atrasentan or a pharmaceuticallyacceptable salt thereof.

In some embodiments, the one or more additional agents are selected fromcalcineurin inhibitors, proteasome inhibitors, aminoquinolines,complement inhibitors, B-cell inhibitors, cytotoxic agents, mTORinhibitors, and steroids.

In certain embodiments, the one or more additional agents are steroids.For example, the one or more additional agents can be selected from thegroup consisting of prednisone, dexamethasone, hydrocortisone,ciclosporin, and combinations of any of the foregoing.

In certain embodiments, the one or more additional agents areaminoquinolines. For example, the one or more additional agents can behydroxychloroquine.

In some embodiments, the subject is receiving one or more additionalagents at the time of treatment with atrasentan. In certain embodiments,the dosage of the one or more additional agents is decreased aftertreatment with atrasentan, or a pharmaceutically acceptable salt thereof(e.g., after 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7weeks, 8 weeks, 9 weeks, 10 weeks, 20 weeks, 30 weeks, 40 weeks, 50weeks, 60 weeks, 70 weeks, 80 weeks, 90 weeks, 100 weeks, 110 weeks, 120weeks, 130 weeks, 140 weeks, 150 weeks, 160 weeks, 170 weeks, 180 weeks,190 weeks, or 200 weeks of treatment). In certain of these embodiments,the dosage of the one or more additional agents is decreased afterbetween about 15 days to about 30 days of treatment with atrasentan, ora pharmaceutically acceptable salt thereof. In certain of the foregoingembodiments, the additional agent dosage is decreased by about 10% toabout 100%. In certain embodiments, the additional agent dosage isdecreased by about 15% to about 100%. In certain embodiments, theadditional agent dosage is decreased by about 20% to about 100%. Incertain embodiments, the additional agent dosage is decreased by about25% to about 100%. In certain embodiments, the additional agent dosageis decreased by about 30% to about 100%. In certain embodiments, theadditional agent dosage is decreased by about 35% to about 100%. Incertain embodiments, the additional agent dosage is decreased by about40% to about 100%. In certain embodiments, the additional agent dosageis decreased by about 45% to about 100%. In certain embodiments, theadditional agent dosage is decreased by about 50% to about 100%. Incertain embodiments, the additional agent dosage is decreased by about55% to about 100%. In certain embodiments, the additional agent dosageis decreased by about 60% to about 100%. In certain embodiments, theadditional agent dosage is decreased by about 65% to about 100%. Incertain embodiments, the additional agent dosage is decreased by about70% to about 100%. In certain embodiments, the additional agent dosageis decreased by about 75% to about 100%. In certain embodiments, theadditional agent dosage is decreased by about 80% to about 100%. Incertain embodiments, the additional agent dosage is decreased by about85% to about 100%. In certain embodiments, the additional agent dosageis decreased by about 90% to about 100%. In certain of the foregoingembodiments, the dosage of the one or more additional agents isdecreased after between about 15 days to about 30 days (e.g., about 15days, about 20 days, about 25 days, or about 30 days) of treatment withatrasentan, or a pharmaceutically acceptable salt thereof. When thedosage of additional agent is decreased by 100% as described herein, thesubject is no longer needing additional agent.

In certain embodiments, the dosage of one or more steroids is decreasedafter treatment with atrasentan, or a pharmaceutically acceptable saltthereof, for example, after between about 15 days to about 30 days oftreatment with atrasentan, or a pharmaceutically acceptable saltthereof. In some embodiments, the steroid dosage is decreased by about10% to about 100%, as described herein. In some embodiments, the dosageof prednisone, dexamethasone, hydrocortisone, ciclosporin, or acombination of any of the foregoing is reduced by about 10% to about100% after treatment with atrasentan, or a pharmaceutically acceptablesalt thereof.

In certain embodiments, the dosage of one or more aminoquinolines isdecreased after treatment with atrasentan, or a pharmaceuticallyacceptable salt thereof, for example, after between about 15 days toabout 30 days of treatment with atrasentan, or a pharmaceuticallyacceptable salt thereof. In some embodiments, the aminoquinoline dosageis decreased by about 10% to about 100%, as described herein. In someembodiments, the dosage of hydroxychloroquine is reduced by about 10% toabout 100% after treatment with atrasentan, or a pharmaceuticallyacceptable salt thereof.

In some embodiments, the subject is concomitantly receiving one or moreadditional therapeutic agents. The one or more additional therapeuticagents are described herein. For example, the subject is concomitantlyreceiving an inhibitor of one or more elements of therenin-angiotensin-aldosterone system. In certain embodiments, thesubject is concomitantly receiving a SGLT-2 inhibitor, an ACE inhibitor,an ARB, a statin, a diuretic, a calcium channel blocker, a beta blocker,an aldosterone antagonist, fish oil, hydroxychloroquine, or acombination of any of the foregoing. In certain of these embodiments,the subject is concomitantly receiving a SGLT-2 inhibitor. In certain ofthese embodiments, the subject is concomitantly receiving an ACEinhibitor, an ARB, or a combination thereof. In certain embodiments, thesubject is concomitantly receiving one or more statins, such asatorvastatin, fluvastatin, lovastatin, pravastatin, rosuvastatin,simvastatin, and pitavastatin. In certain embodiments, the subject isconcomitantly receiving one or more diuretics, such ashydrochlorothiazide, trichlormethiazide, hydroflumethiazide,quinethazone, metolazone, chlorothiazide, chlorthalidone, indapamide,methyclothiazide bemetanide, torsemide, piretanide, ethacrynic acid,bumetanide, furosemide, triamterene, spironolactone, eplerenone, andamiloride. In certain embodiments, the subject is concomitantlyreceiving a SGLT-2 inhibitor, such as canagliflozin, dapagliflozin,empagliflozin, or ertugliflozin. In certain embodiments, the subject isconcomitantly receiving one or more ACE inhibitors, such as quinapril,fosinopril perindopril, captopril, enalapril, enalaprilat, ramipril,cilazapril, delapril, fosenopril, zofenopril, indolapril, benazepril,lisinopril, spirapril, trandolapril, perindep, pentopril, moexipril,rescinnamine, and pivopril. In certain embodiments, the subject isconcomitantly receiving an ARB, such as candesartan, candesartancilexetil, eprosartan, irbesartan, losartan, olmesartan, olmesartanmedoxomil, telmisartan, valsartan, azilsartan medoxomil, and BRA-657. Incertain embodiments, the subject is concomitantly receiving a diureticand an ACE inhibitor or an ARB. In certain embodiments, the subject isconcomitantly receiving a diuretic, an ACE inhibitor, and an ARB. Incertain embodiments, the subject is concomitantly receiving a diureticand a SGLT-2 inhibitor, and an ACE inhibitor or an ARB. In certainembodiments, the subject is concomitantly receiving a diuretic, a SGLT-2inhibitor, an ACE inhibitor, and an ARB. In certain embodiments, thesubject concomitantly receiving one or more additional therapeuticagents has not previously received the one or more therapeutic agents.For example, a subject that is concomitantly receiving a SGLT-2inhibitor that has not previously received a SGLT-2 inhibitor.

In some embodiments, the subject has previously received, but is notconcomitantly receiving, one or more additional therapeutic agents suchas those described herein. For example, the subject is has previouslyreceived, but is not concomitantly receiving a SGLT-2 inhibitor, an ACEinhibitor, an ARB, a statin, a diuretic, a calcium channel blocker, abeta blocker, an aldosterone antagonist, fish oil, hydroxychloroquine,or a combination of any of the foregoing, as described herein. Incertain of these embodiments, the subject has previously received, butis not concomitantly receiving a SGLT-2 inhibitor.

In some embodiments, the subject has cellular glomerular crescentspresent in about ≤25% of glomeruli within 6 months prior to the firstadministration of atrasentan or a pharmaceutically acceptable saltthereof. For example, the subject can have cellular glomerular crescentspresent in about 25%, about 20%, about 15%, about 10%, about 5%, orabout 1%, or any value in between, of glomeruli. In some embodiments,the subject does not have cellular glomerular crescents present in theglomeruli. In certain embodiments, the subject is not under clinicalsuspicion of rapidly progressive glomerulonephritis (RPGN).

In some embodiments, the subject has not undergone organ transplantationprior to the first administration of atrasentan or a pharmaceuticallyacceptable salt thereof.

In some embodiments, the subject has a systolic blood pressure of belowabout 160 mmHg prior to the first administration of atrasentan or apharmaceutically acceptable salt thereof. For example, the subject canbe a systolic blood pressure of below about 155 mmHg, below about 150mmHg, below about 145 mmHg, or below about 140 mmHg. In someembodiments, the subject has a diastolic blood pressure of below about100 mmHg prior to the first administration of atrasentan or apharmaceutically acceptable salt thereof. For example, the subject canhave a diastolic blood pressure of below about 100 mmHg, below about 95mmHg, or below about 90 mmHg. In some embodiments, the subject has asystolic blood pressure of between about 100 mm Hg and about 130 mm Hgand a diastolic blood pressure of about 70 mm Hg to about 90 mm Hg.

In some embodiments, the subject has not been diagnosed with heartfailure prior to the first administration of atrasentan or apharmaceutically acceptable salt thereof. In some embodiments, thesubject has not been previously admitted to hospital for conditionsrelating to fluid overload prior to the first administration ofatrasentan or a pharmaceutically acceptable salt thereof. Non-limitingexamples of conditions include uncontrolled peripheral edema, pleuraleffusion, or ascites. In some embodiments, the subject has not beendiagnosed with clinically significant liver disease prior to the firstadministration of atrasentan or a pharmaceutically acceptable saltthereof. In some embodiments, the transaminase or bilirubin values ofthe subject are no more than twice the normal upper limit prior to thefirst administration of atrasentan or a pharmaceutically acceptable saltthereof. For example, the ALT level of the subject is below about 110U/L (e.g., below about 100 U/L, below 90 U/L, below about 80 U/L, belowabout 70 U/L, below about 60 U/L, below about 50 U/L, or below about 40U/L, or any value in between). As another example, the AST level of thesubject is below 100 U/L (e.g., below 90 U/L, below about 80 U/L, belowabout 70 U/L, below about 60 U/L, below about 50 U/L, or below about 40U/L, or any value in between). As yet another example, the bilirubinlevel of the subject is below about 2.5 mg/dL (e.g., below about 2mg/dL, below about 1.5 mg/dL, below about 1.4 mg/dL, below about 1.3mg/dL, below about 1.2 mg/dL, below about 1.1 mg/dL, below about 1.0mg/dL, or below about 0.9 mg/dL, or any value in between).

In some embodiments, the subject has a hemoglobin level of above about 9g/dL (e.g., above about 10 g/dL, about 11 g/dL, about 12 g/dL, or about13 g/dL, or any value in between) prior to the first administration ofatrasentan or a pharmaceutically acceptable salt thereof. In someembodiments, the subject has not received blood transfusion for anemiafor at least about 3 months (e.g., at least about 4 months, about 5months, about 6 months, or about one year) prior to the firstadministration of atrasentan or a pharmaceutically acceptable saltthereof. In some embodiments, the subject has not been diagnosed withcancer for at least 5 years prior to the first administration ofatrasentan or a pharmaceutically acceptable salt thereof. In someembodiments, the subject has not been diagnosed with cancer (e.g., lungcancer or prostate cancer) for at least 5 years prior to the firstadministration of atrasentan or a pharmaceutically acceptable saltthereof. In some embodiments, the subject has not been diagnosed withcancer for at least 5 years prior to the first administration ofatrasentan or a pharmaceutically acceptable salt thereof, unless thecancer is nonmelanoma skin cancer not requiring ongoing treatment. Insome embodiments, the subject does not have cancer prior to the firstadministration of atrasentan or a pharmaceutically acceptable saltthereof, unless the cancer is nonmelanoma skin cancer not requiringongoing treatment. In some embodiments, the subject does not suffer fromcancer prior to the first administration of atrasentan or apharmaceutically acceptable salt thereof, unless the cancer isnonmelanoma skin cancer not requiring ongoing treatment. In someembodiments, the subject is not being treated for cancer for at least 5years prior to the first administration of atrasentan or apharmaceutically acceptable salt thereof, unless the cancer isnonmelanoma skin cancer not requiring ongoing treatment.

In some embodiments of the methods, uses, or product for uses herein,the subject has not been previously diagnosed with one or more ofdiabetic nephropathy, HIV/AIDS, or acute kidney failure. In someembodiments of the methods, uses, or product for uses herein, thesubject has not been previously diagnosed with one or more of diabeticnephropathy, HIV/AIDS, cancer (e.g., prostate cancer or lung cancer), oracute kidney failure. In some embodiments, the subject has not beenpreviously diagnosed with one or more of diabetic nephropathy, HIV/AIDS,prostate cancer, or acute kidney failure. In some embodiments, thesubject has not been previously diagnosed with one or more of diabeticnephropathy, HIV-related nephropathy, prostate cancer, or acute kidneyfailure. In some embodiments, the subject has not been previouslydiagnosed with one or more of diabetic nephropathy, HIV-relatednephropathy, cancer (e.g., lung cancer, or prostate cancer), or acutekidney failure. In some embodiments, the subject has not been previouslydiagnosed with one or more of diabetic nephropathy, HIV-relatednephropathy, or acute kidney failure. In certain embodiments, thesubject has not been previously diagnosed with diabetic nephropathy. Incertain embodiments, the subject has not been previously diagnosed withHIV/AIDS. In certain embodiments, the subject has not been previouslydiagnosed with acute kidney failure. In certain embodiments, the subjecthas not been previously diagnosed with HIV-related nephropathy. Incertain embodiments, the subject has not been diagnosed with cancer. Incertain embodiments, the subject has not been diagnosed with prostatecancer. In certain embodiments, the subject has not been diagnosed withlung cancer. In certain embodiments, the subject has not been previouslydiagnosed with any one of diabetic nephropathy, HIV/AIDS, and acutekidney failure. In certain embodiments, the subject has not beenpreviously diagnosed with any one of diabetic nephropathy, HIV/AIDS,prostate cancer, and acute kidney failure. In certain embodiments, thesubject has not been previously diagnosed with any one of diabeticnephropathy, HIV-related nephropathy, prostate cancer, and acute kidneyfailure. In certain embodiments, the subject has not been previouslydiagnosed with any one of diabetic nephropathy, HIV-related nephropathy,and acute kidney failure. In some embodiments of the methods, uses, orproduct for uses herein, the subject has not been previously diagnosedwith diabetes. In some embodiments of the methods, uses, or product foruses herein, the subject has not been previously diagnosed with Type 2diabetes. In certain of the foregoing embodiments, the subject has beendetermined to have controlled serum glucose levels as described anywhereherein.

In some embodiments, the subject does not have one or more of diabeticnephropathy, HIV/AIDS, or acute kidney failure. In some embodiments, thesubject does not have one or more of diabetic nephropathy, HIV/AIDS,prostate cancer, or acute kidney failure. In some embodiments, thesubject does not have one or more of diabetic nephropathy, HIV-relatednephropathy, prostate cancer, or acute kidney failure. In someembodiments, the subject does not have one or more of diabeticnephropathy, HIV-related nephropathy, or acute kidney failure. Incertain embodiments, the subject does not have diabetic nephropathy. Incertain embodiments, the subject does not have HIV/AIDS. In certainembodiments, the subject does not have acute kidney failure. In certainembodiments, the subject does not have HIV-related nephropathy. Incertain embodiments, the subject does not have prostate cancer. Incertain embodiments, the subject does not have any one of diabeticnephropathy, HIV/AIDS, and acute kidney failure. In certain embodiments,the subject does not have any one of diabetic nephropathy, HIV/AIDS,prostate cancer, and acute kidney failure. In certain embodiments, thesubject does not have any one of diabetic nephropathy, HIV-relatednephropathy, prostate cancer, and acute kidney failure. In certainembodiments, the subject does not have any one of diabetic nephropathy,HIV-related nephropathy, and acute kidney failure. In some embodiments,the subject does not have diabetes. In some embodiments, the subjectdoes not have Type 2 diabetes. In certain of the foregoing embodiments,the subject has been determined to have controlled serum glucose levelsas described anywhere herein.

In some embodiments, the subject does not suffer from one or more ofdiabetic nephropathy, HIV/AIDS, or acute kidney failure. In someembodiments, the subject does not suffer from one or more of diabeticnephropathy, HIV/AIDS, prostate cancer, or acute kidney failure. In someembodiments, the subject does not suffer from one or more of diabeticnephropathy, HIV-related nephropathy, prostate cancer, or acute kidneyfailure. In some embodiments, the subject does not suffer from one ormore of diabetic nephropathy, HIV-related nephropathy, or acute kidneyfailure. In certain embodiments, the subject does not suffer fromdiabetic nephropathy. In certain embodiments, the subject does notsuffer from HIV/AIDS. In certain embodiments, the subject does notsuffer from acute kidney failure. In certain embodiments, the subjectdoes not suffer from HIV-related nephropathy. In certain embodiments,the subject does not suffer from prostate cancer. In certainembodiments, the subject does not suffer from any one of diabeticnephropathy, HIV/AIDS, and acute kidney failure. In certain embodiments,the subject does not suffer from any one of diabetic nephropathy,HIV/AIDS, prostate cancer, and acute kidney failure. In certainembodiments, the subject does not suffer from any one of diabeticnephropathy, HIV-related nephropathy, prostate cancer, and acute kidneyfailure. In certain embodiments, the subject does not suffer from anyone of diabetic nephropathy, HIV-related nephropathy, and acute kidneyfailure. In some embodiments, the subject does not suffer from diabetes.In some embodiments, the subject does not suffer from Type 2 diabetes.In certain of the foregoing embodiments, the subject has been determinedto have controlled serum glucose levels as described anywhere herein.

In some embodiments, the subject is not being treated for one or more ofdiabetic nephropathy, HIV/AIDS, or acute kidney failure. In someembodiments, the subject is not being treated for one or more ofdiabetic nephropathy, HIV/AIDS, prostate cancer, or acute kidneyfailure. In some embodiments, the subject is not being treated for oneor more of diabetic nephropathy, HIV-related nephropathy, prostatecancer, or acute kidney failure. In some embodiments, the subject is notbeing treated for one or more of diabetic nephropathy, HIV-relatednephropathy, or acute kidney failure. In certain embodiments, thesubject is not being treated for diabetic nephropathy. In certainembodiments, the subject is not being treated for HIV/AIDS. In certainembodiments, the subject is not being treated for acute kidney failure.In certain embodiments, the subject is not being treated for HIV-relatednephropathy. In certain embodiments, the subject is not being treatedfor prostate cancer. In certain embodiments, the subject is not beingtreated for any one of diabetic nephropathy, HIV/AIDS, and acute kidneyfailure. In certain embodiments, the subject is not being treated forany one of diabetic nephropathy, HIV/AIDS, prostate cancer, and acutekidney failure. In certain embodiments, the subject is not being treatedfor any one of diabetic nephropathy, HIV-related nephropathy, prostatecancer, and acute kidney failure. In certain embodiments, the subject isnot being treated for any one of diabetic nephropathy, HIV-relatednephropathy, and acute kidney failure. In some embodiments, the subjectis not being treated for diabetes. In some embodiments, the subject isnot being treated for Type 2 diabetes. In certain of the foregoingembodiments, the subject has been determined to have controlled serumglucose levels as described anywhere herein.

In some embodiments, the subject has been determined to have controlledserum glucose levels; or the subject has not been diagnosed with one ormore of HIV-related nephropathy or acute kidney failure. In certainembodiments, the subject has been determined to have controlled serumglucose levels. For example, the subject has been determined to have afasting serum glucose level of below about 130 mg/dL, about 125 mg/dL,about 120 mg/dL, about 115 mg/dL, about 110 mg/dL, about 105 mg/dL,about 100 mg/dL, about 95 mg/dL, about 90 mg/dL, about 85 mg/dL, about80 mg/dL, or about 75 mg/dL, or any value in between. In certainembodiments, the subject has not been diagnosed with one or more ofHIV-related nephropathy or acute kidney failure. In certain embodiments,the subject has been determined to have controlled serum glucose levelsas described anywhere herein; and the subject has not been diagnosedwith one or more of HIV-related nephropathy or acute kidney failure.

In some embodiments, the subject has not been previously diagnosed witha chronic kidney disease that is other than IgA nephropathy.Non-limiting examples include a diabetic kidney disease, a hypertensivekidney disease, or a primary glomerulopathy that is determined to not beassociated with IgA nephropathy. In certain embodiments, the subject hasnot been previously diagnosed with a diabetic kidney disease. In certainembodiments, the subject has not been previously diagnosed with ahypertensive kidney disease. In certain embodiments, the subject has notbeen diagnosed with a primary glomerulopathy that is determined to notbe associated with IgA nephropathy.

In some embodiments, the subject does not have a chronic kidney diseasethat is other than IgA nephropathy. Non-limiting examples include adiabetic kidney disease, a hypertensive kidney disease, or a primaryglomerulopathy that is determined to not be associated with IgAnephropathy. In certain embodiments, the subject does not have adiabetic kidney disease. In certain embodiments, the subject does nothave a hypertensive kidney disease. In certain embodiments, the subjectdoes not have a primary glomerulopathy that is determined to not beassociated with IgA nephropathy.

In some embodiments, the subject does not suffer from a chronic kidneydisease that is other than IgA nephropathy. Non-limiting examplesinclude a diabetic kidney disease, a hypertensive kidney disease, or aprimary glomerulopathy that is determined to not be associated with IgAnephropathy. In certain embodiments, the subject does not suffer from adiabetic kidney disease. In certain embodiments, the subject does notsuffer from a hypertensive kidney disease. In certain embodiments, thesubject does not suffer from a primary glomerulopathy that is determinedto not be associated with IgA nephropathy.

In some embodiments, the subject is not being treated for a chronickidney disease that is other than IgA nephropathy. Non-limiting examplesinclude a diabetic kidney disease, a hypertensive kidney disease, or aprimary glomerulopathy that is determined to not be associated with IgAnephropathy. In certain embodiments, the subject is not being treatedfor a diabetic kidney disease. In certain embodiments, the subject isnot being treated for a hypertensive kidney disease. In certainembodiments, the subject is not being treated for a primaryglomerulopathy that is determined to not be associated with IgAnephropathy.

Treatment Outcome

In some embodiments of the methods, uses, or product for uses herein,renal inflammation is decreased after treatment with atrasentan or apharmaceutically acceptable salt thereof. In some embodiments, renalinflammation in the subject is decreased by at least about 10% aftertreatment with atrasentan or a pharmaceutically acceptable salt thereof(e.g., after treatment for 1 week, about 2 weeks, about 3 weeks, about 4weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about9 weeks, about 10 weeks, about 20 weeks, about 30 weeks, about 40 weeks,about 50 weeks, about 60 weeks, about 70 weeks, about 80 weeks, about 90weeks, about 100 weeks, about 110 weeks, about 120 weeks, about 130weeks, about 140 weeks, about 150 weeks, about 160 weeks, about 170weeks, about 180 weeks, about 190 weeks, or about 200 weeks, or anyvalue in between). In some embodiments, the renal inflammation in thesubject is decreased by at least about 20%, about 30%, about 40%, about50%, about 60% about 70% about 80%, about 90%, or about 95%, or anyvalue in between. In certain of the foregoing embodiments, the subjecthas been treated with atrasentan or a pharmaceutically acceptable saltthereof for between about 15 days and about 30 days.

In some embodiments, renal fibrosis is decreased after treatment withatrasentan or a pharmaceutically acceptable salt thereof. In someembodiments, renal fibrosis in the subject is decreased by at leastabout 10% after treatment with atrasentan or a pharmaceuticallyacceptable salt thereof (e.g., after treatment for 1 week, about 2weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 20 weeks,about 30 weeks, about 40 weeks, about 50 weeks, about 60 weeks, about 70weeks, about 80 weeks, about 90 weeks, about 100 weeks, about 110 weeks,about 120 weeks, about 130 weeks, about 140 weeks, about 150 weeks,about 160 weeks, about 170 weeks, about 180 weeks, about 190 weeks, orabout 200 weeks, or any value in between). In certain embodiments, therenal fibrosis in the subject is decreased by at least about 20%, about30%, about 40%, about 50%, about 60% about 70% about 80%, about 90%, orabout 95%, or any value in between. In certain of the foregoingembodiments, the subject has been treated with atrasentan or apharmaceutically acceptable salt thereof for between about 15 days andabout 30 days.

In some embodiments, renal fibrosis in the subject is decreased to lessthan about 50% of the cortical area after treatment with atrasentan or apharmaceutically acceptable salt thereof (e.g., after treatment for 1week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks,about 20 weeks, about 30 weeks, about 40 weeks, about 50 weeks, about 60weeks, about 70 weeks, about 80 weeks, about 90 weeks, about 100 weeks,about 110 weeks, about 120 weeks, about 130 weeks, about 140 weeks,about 150 weeks, about 160 weeks, about 170 weeks, about 180 weeks,about 190 weeks, or about 200 weeks, or any value in between). Incertain embodiments, renal fibrosis in the subject is decreased to lessthan about 40% of the cortical area. For example, in some embodiments,renal fibrosis in the subject is decreased to less than about 35%, about30%, about 25%, about 20%, about 15%, or about 10%, or any value inbetween, of the cortical area. In certain of the foregoing embodiments,the subject has been treated with atrasentan or a pharmaceuticallyacceptable salt thereof for between about 15 days and about 30 days.

In some embodiments, the occurrence of hematuria is decreased in asubject after treatment with atrasentan or a pharmaceutically acceptablesalt thereof. In some embodiments, the number of urinary red blood cellsper high powered (microscope) field (rbc/hpf) in the subject isdecreased by at least about 10% after treatment with atrasentan or apharmaceutically acceptable salt thereof (e.g., after treatment for 1week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks,about 20 weeks, about 30 weeks, about 40 weeks, about 50 weeks, about 60weeks, about 70 weeks, about 80 weeks, about 90 weeks, about 100 weeks,about 110 weeks, about 120 weeks, about 130 weeks, about 140 weeks,about 150 weeks, about 160 weeks, about 170 weeks, about 180 weeks,about 190 weeks, or about 200 weeks). In certain embodiments, theurinary rbc/hpf in the subject is decreased by at least about 20%. Forexample, in some embodiments, the urinary rbc/hpf in the subject isdecreased by at least about 30%, about 40%, about 50%, about 60% about70% about 80%, about 90%, or about 95%, or any value in between. Incertain of the foregoing embodiments, the subject has been treated withatrasentan or a pharmaceutically acceptable salt thereof for betweenabout 15 days and about 30 days.

In some embodiments, the rate of decrease of eGFR of the subject isreduced by at least about 10% after treatment with atrasentan or apharmaceutically acceptable salt thereof (e.g., after treatment for 1week, 2 weeks, 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks,about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 20weeks, about 30 weeks, about 40 weeks, about 50 weeks, about 60 weeks,about 70 weeks, about 80 weeks, about 90 weeks, about 100 weeks, about110 weeks, about 120 weeks, about 130 weeks, about 140 weeks, about 150weeks, about 160 weeks, about 170 weeks, about 180 weeks, about 190weeks, or about 200 weeks, or any value in between). In certainembodiments, the rate of decrease of eGFR of the subject is reduced byat least about 20%. For example, in some embodiments, the rate ofdecrease of eGFR of the subject is reduced by at least about 30%, about40%, about 50%, about 60% about 70% about 80%, about 90%, or about 95%,or any value in between. In certain of the foregoing embodiments, thesubject has been treated with atrasentan or a pharmaceuticallyacceptable salt thereof for between about 15 days and about 30 days. Incertain of the foregoing embodiments, the subject has been treated withatrasentan or a pharmaceutically acceptable salt thereof for betweenabout 6 months and about 1 year.

In some embodiments, the rate of decrease of eGFR of the subject isreduced to below about 10 mL/min per year after treatment withatrasentan or a pharmaceutically acceptable salt thereof (e.g., aftertreatment for about 1 week, about 2 weeks, about 3 weeks, about 4 weeks,about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9weeks, about 10 weeks, about 20 weeks, about 30 weeks, about 40 weeks,about 50 weeks, about 60 weeks, about 70 weeks, about 80 weeks, about 90weeks, about 100 weeks, about 110 weeks, about 120 weeks, about 130weeks, about 140 weeks, about 150 weeks, about 160 weeks, about 170weeks, about 180 weeks, about 190 weeks, or about 200 weeks, or anyvalue in between). In some embodiments, the rate of decrease of eGFR ofthe subject is reduced to below about 9 mL/min per year. For example, insome embodiments, the rate of decrease of eGFR of the subject is reducedto below about 8 mL/min per year, about 7 mL/min per year, about 6mL/min per year, about 5 mL/min per year, about 4 mL/min per year, about3 mL/min per year, about 2 mL/min per year, or about 1 mL/min per year,or any value in between. In certain of the foregoing embodiments, thesubject has been treated with atrasentan or a pharmaceuticallyacceptable salt thereof for between about 15 days and about 30 days. Incertain of the foregoing embodiments, the subject has been treated withatrasentan or a pharmaceutically acceptable salt thereof for betweenabout 6 months and about 1 year.

In some embodiments, the risk of the subject developing ESRD is reducedby about 20% to about 99% after treatment with atrasentan or apharmaceutically acceptable salt thereof (e.g., after treatment for 1week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks,about 20 weeks, about 30 weeks, about 40 weeks, about 50 weeks, about 60weeks, about 70 weeks, about 80 weeks, about 90 weeks, about 100 weeks,about 110 weeks, about 120 weeks, about 130 weeks, about 140 weeks,about 150 weeks, about 160 weeks, about 170 weeks, about 180 weeks,about 190 weeks, or about 200 weeks, or any value in between). Forexample, the risk of the subject developing ESRD can be reduced by about20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%,about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about85%, about 90%, or about 99%, or any value in between. In certain of theforegoing embodiments, the subject has been treated for between about 90days to about 180 days. In certain embodiments, the risk of the subjectdeveloping ESRD is reduced by about 20% to about 99% after between about90 and about 180 days of treatment with atrasentan, or apharmaceutically acceptable salt thereof. In certain of the foregoingembodiments, the subject has been treated with atrasentan or apharmaceutically acceptable salt thereof for between about 6 months andabout 1 year.

In some embodiments, the method increases the time between the diagnosisof IgA nephropathy in the subject and the time when eGFR of the subjectfalls below about 15 mL/min/1.73 m². In certain embodiments, the methodincreases the time between the diagnosis of IgA nephropathy in thesubject and the time when eGFR of the subject falls below 15 mL/min/1.73m² by at least about 10%. For example, in some embodiments, the methodincreases the time between the diagnosis of IgA nephropathy in thesubject and the time when eGFR of the subject falls below about 15mL/min/1.73 m² by at least about 20%, about 30%, about 40%, about 50%,about 60%, about 70%, about 80%, about 90%, about 95%, about 100%, about150%, about 200%, about 250%, about 300%, about 350%, about 400%, about450%, or about 500%, or any value in between.

In some embodiments, the method increases the time between the diagnosisof IgA nephropathy in the subject and the time when eGFR of the subjectfalls below 15 mL/min/1.73 m² by at least about 1 year. For example, themethod can delay the time when eGFR of the subject falls below 15mL/min/1.73 m² by at least about 1.5 years, about 2 years, about 2.5years, about 3 years, about 3.5 years, about 4 years, about 4.5 years,about 5 years, about 5.5 years, about 6 years, about 6.5 years, about 7years, about 7.5 years, about 8 years, about 8.5 years, about 9 years,about 9.5 years, about 10 years, about 11 years, about 12 years, about13 years, about 15 years, about 15 years, about 16 years, about 17years, about 18 years, about 19 years, or about 20 years, or any valuein between.

In some embodiments, the method reduces the average rate of decrease ineGFR by from about 0.75 mL/min/year to about 6 mL/min/year for at leastabout 3 months (e.g., at least about 4 months, at least about 5 months,at least about 6 months, at least about 7 months, at least about 8months, at least about 9 months, at least about 10 months, at leastabout 11 months, at least about a year, at least about 1.5 years, or atleast about 2 years) prior to the first administration of atrasentan, ora pharmaceutically acceptable salt thereof. For example, the methodreduces the average rate of decrease in eGFR by about 0.75 mL/min/year,about 1 mL/min/year, about 1.5 mL/min/year, about 2 mL/min/year, about2.5 mL/min/year, about 3 mL/min/year, about 3.5 mL/min/year, about 4mL/min/year, about 4.5 mL/min/year, about 5 mL/min/year, about 5.5mL/min/year, or about 6 mL/min/year. In some embodiments, the methodreduces the average rate of decrease in eGFR by from about 4 mL/min/yearto about 5 mL/min/year for at least about 3 months prior to the firstadministration of atrasentan, or a pharmaceutically acceptable saltthereof. In some embodiments, the method reduces the average rate ofdecrease in eGFR by from about 3 mL/min/year to about 6 mL/min/year forat least about 3 months prior to the first administration of atrasentan,or a pharmaceutically acceptable salt thereof. In some embodiments, themethod reduces the average rate of decrease in eGFR by from about 4mL/min/year to about 5 mL/min/year for at least about 3 months prior tothe first administration of atrasentan, or a pharmaceutically acceptablesalt thereof. In some embodiments, the decrease in eGFR in mL/min/yearrefers to units per 1.73 m².

In some embodiments, the method reduces the average rate of decrease ineGFR by from about 15% to about 30% after between about 6 months andabout 24 months of treatment with atrasentan, or a pharmaceuticallyacceptable salt thereof. In some embodiments, the average rate ofdecrease in eGFR may be reduced by about 15%, after about 6 months, 9,months, 12 months, 15 months, 18 months, 21 months, or 24 months oftreatment with atrasentan, or a pharmaceutically acceptable saltthereof. In some embodiments, the average rate of decrease in eGFR maybe reduced by about 20%, after about 6 months, 9, months, 12 months, 15months, 18 months, 21 months, or 24 months of treatment with atrasentan,or a pharmaceutically acceptable salt thereof. In some embodiments, theaverage rate of decrease in eGFR may be reduced by about 25%, afterabout 6 months, 9, months, 12 months, 15 months, 18 months, 21 months,or 24 months of treatment with atrasentan, or a pharmaceuticallyacceptable salt thereof. In some embodiments, the average rate ofdecrease in eGFR may be reduced by about 30%, after about 6 months, 9,months, 12 months, 15 months, 18 months, 21 months, or 24 months oftreatment with atrasentan, or a pharmaceutically acceptable saltthereof.

In another aspect, provided herein is a method of decreasingproteinuria, comprising administering a therapeutically effective amountof atrasentan, or a pharmaceutically acceptable salt thereof, to asubject in need thereof.

In some embodiments, the amount of proteins (e.g., albumin) in the urineof the subject is reduced by at least about 10% after treatment withatrasentan or a pharmaceutically acceptable salt thereof (e.g., aftertreatment for about 1 week, about 2 weeks, about 3 weeks, about 4 weeks,about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9weeks, about 10 weeks, about 20 weeks, about 30 weeks, about 40 weeks,about 50 weeks, about 60 weeks, about 70 weeks, about 80 weeks, about 90weeks, about 100 weeks, about 110 weeks, about 120 weeks, about 130weeks, about 140 weeks, about 150 weeks, about 160 weeks, about 170weeks, about 180 weeks, about 190 weeks, or about 200 weeks, or anyvalue in between). In some embodiments, the amount of proteins in theurine of the subject is reduced by at least about 15%. For example, insome embodiments, the amount of proteins in the urine of the subject isreduced by at least about 20%, about 30%, about 40%, about 50%, about60%, about 70%, about 80%, about 90%, or about 95%, or any value inbetween. In certain of the foregoing embodiments, the subject has beentreated with atrasentan or a pharmaceutically acceptable salt thereoffor between about 15 days and about 30 days.

In certain embodiments, the amount of proteins (e.g., albumin) in theurine of the subject is reduced by about 20% to about 80% after betweenabout 15 day and about 30 days of treatment with atrasentan, or apharmaceutically acceptable salt thereof. In certain of theseembodiments, the amount of proteins in the urine of the subject isreduced by about 25% to about 80%. In certain of these embodiments, theamount of proteins in the urine of the subject is reduced by about 30%to about 80%. In certain of these embodiments, the amount of proteins inthe urine of the subject is reduced by about 35% to about 80%. Incertain of these embodiments, the amount of proteins in the urine of thesubject is reduced by about 40% to about 80%. In certain of theseembodiments, the amount of proteins in the urine of the subject isreduced by about 45% to about 80%. In certain of these embodiments, theamount of proteins in the urine of the subject is reduced by about 50%to about 80%.

In some embodiments, the amount of proteins (e.g., albumin) in the urineof the subject is reduced by about 100 mg/dL to about 3,000 mg/dL aftertreatment with atrasentan or a pharmaceutically acceptable salt thereof(e.g., after treatment for about 1 week, about 2 weeks, about 3 weeks,about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8weeks, about 9 weeks, about 10 weeks, about 20 weeks, about 30 weeks,about 40 weeks, about 50 weeks, about 60 weeks, about 70 weeks, about 80weeks, about 90 weeks, about 100 weeks, about 110 weeks, about 120weeks, about 130 weeks, about 140 weeks, about 150 weeks, about 160weeks, about 170 weeks, about 180 weeks, about 190 weeks, or about 200weeks, or any value in between). In certain embodiments, the amount ofproteins in the urine of the subject is reduced by about 100 mg/dL toabout 2,500 mg/dL. In certain embodiments, the amount of proteins in theurine of the subject is reduced by about 100 mg/dL to about 2,000 mg/dL.In certain embodiments, the amount of proteins in the urine of thesubject is reduced by about 100 mg/dL to about 1,500 mg/dL. In certainembodiments, the amount of proteins in the urine of the subject isreduced by about 100 mg/dL to about 1,000 mg/dL. In certain embodiments,the amount of proteins in the urine of the subject is reduced by about100 mg/dL to about 500 mg/dL. In certain embodiments, the amount ofproteins in the urine of the subject is reduced by about 100 mg/dL toabout 400 mg/dL. In certain embodiments, the amount of proteins in theurine of the subject is reduced by about 100 mg/dL to about 300 mg/dL.In certain embodiments, the amount of proteins in the urine of thesubject is reduced by about 100 mg/dL to about 200 mg/dL. In certainembodiments, the amount of proteins in the urine of the subject isreduced by about 500 mg/dL to about 2,500 mg/dL. In certain embodiments,the amount of proteins in the urine of the subject is reduced by about500 mg/dL to about 2,000 mg/dL. In certain embodiments, the amount ofproteins in the urine of the subject is reduced by about 500 mg/dL toabout 1,500 mg/dL. In certain embodiments, the amount of proteins in theurine of the subject is reduced by about 500 mg/dL to about 1,000 mg/dL.In certain embodiments, the amount of proteins in the urine of thesubject is reduced by about 500 mg/dL to about 900 mg/dL. In certainembodiments, the amount of proteins in the urine of the subject isreduced by about 500 mg/dL to about 800 mg/dL. In certain embodiments,the amount of proteins in the urine of the subject is reduced by about600 mg/dL to about 900 mg/dL. In certain embodiments, the amount ofproteins in the urine of the subject is reduced by about 700 mg/dL toabout 900 mg/dL. In certain embodiments, the amount of proteins in theurine of the subject is reduced by about 1,000 mg/dL to about 2,000mg/dL. In certain of the foregoing embodiments, the subject has beentreated with atrasentan or a pharmaceutically acceptable salt thereoffor between about 15 days and about 30 days.

In certain embodiments, the amount of proteins (e.g., albumin) in theurine of the subject is reduced by about 100 mg/dL to about 500 mg/dLafter between about 15 days and about 30 days of treatment withatrasentan, or a pharmaceutically acceptable salt thereof. In certainembodiments, the amount of proteins in the urine of the subject isreduced by about 200 mg/dL to about 500 mg/dL after between about 15days and about 30 days of treatment with atrasentan, or apharmaceutically acceptable salt thereof. In certain embodiments, theamount of proteins in the urine of the subject is reduced by about 300mg/dL to about 500 mg/dL after between about 15 days and about 30 daysof treatment with atrasentan, or a pharmaceutically acceptable saltthereof.

In certain embodiments, the amount of proteins (e.g., albumin) in theurine of the subject is reduced by about 500 mg/dL to about 900 mg/dLafter between about 15 days and about 30 days of treatment withatrasentan, or a pharmaceutically acceptable salt thereof. In certainembodiments, the amount of proteins in the urine of the subject isreduced by about 600 mg/dL to about 900 mg/dL after between about 15days and about 30 days of treatment with atrasentan, or apharmaceutically acceptable salt thereof. In certain embodiments, theamount of proteins in the urine of the subject is reduced by about 700mg/dL to about 900 mg/dL after between about 15 days and about 30 daysof treatment with atrasentan, or a pharmaceutically acceptable saltthereof.

In some embodiments, the subject has a reduced level of proteins (e.g.,albumin) in the urine of below about 1.0 gram/day after treatment withatrasentan or a pharmaceutically acceptable salt thereof (e.g., aftertreatment for about 1 week, about 2 weeks, about 3 weeks, about 4 weeks,about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9weeks, about 10 weeks, about 20 weeks, about 30 weeks, about 40 weeks,about 50 weeks, about 60 weeks, about 70 weeks, about 80 weeks, about 90weeks, about 100 weeks, about 110 weeks, about 120 weeks, about 130weeks, about 140 weeks, about 150 weeks, about 160 weeks, about 170weeks, about 180 weeks, about 190 weeks, or about 200 weeks). In someembodiments, the subject has a reduced level of proteins in the urine ofbelow about 0.9 gram/day. For example, in some embodiments, the subjecthas a reduced level of proteins in the urine of below about 0.8gram/day, about 0.7 gram/day, about 0.6 gram/day, 0.5 gram/day, about0.4 gram/day, about 0.3 gram/day, or about 0.2 gram/day, or any value inbetween. In certain of the foregoing embodiments, the subject has beentreated with atrasentan or a pharmaceutically acceptable salt thereoffor between about 15 days and about 30 days.

In some embodiments, the subject is between about 15 and about 40 yearsold. In some embodiments, the subject is between about 15 to about 25years old, about 20 to about 30 years old, about 25 to about 35 yearsold, about 30 to about 40 years old, or any age in between. In someembodiments, the subject is between about 20 to about 30 years old, orany age in between. In some embodiments, the subject is about 20 yearsold, about 21 years old, about 22 years old, about 23 years old, about24 years old, about 25 years old, about 26 years old, about 27 yearsold, about 28 years old, about 29 years old, or about 30 years old.

In some embodiments, the level of fatigue of the patient is reducedfollowing treatment with atrasentan or a pharmaceutically acceptablesalt thereof. In some embodiments, the fatigue is reduced by about 5% toabout 80% after treatment with atrasentan or a pharmaceuticallyacceptable salt thereof (e.g., after treatment for about 1 week, about 2weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 20 weeks,about 30 weeks, about 40 weeks, about 50 weeks, about 60 weeks, about 70weeks, about 80 weeks, about 90 weeks, about 100 weeks, about 110 weeks,about 120 weeks, about 130 weeks, about 140 weeks, about 150 weeks,about 160 weeks, about 170 weeks, about 180 weeks, about 190 weeks, orabout 200 weeks, or any value in between). In certain embodiments, thefatigue is reduced by about 10% to about 75%. In certain embodiments,the fatigue is reduced by about 10% to about 70%. In certainembodiments, the fatigue is reduced by about 10% to about 65%. Incertain embodiments, the fatigue is reduced by about 10% to about 60%.In certain embodiments, the fatigue is reduced by about 10% to about55%. In certain embodiments, the fatigue is reduced by about 10% toabout 50%. In certain embodiments, the fatigue is reduced by about 10%to about 45%. In certain embodiments, the fatigue is reduced by about10% to about 40%. In certain embodiments, the fatigue is reduced byabout 10% to about 35%. In certain embodiments, the fatigue is reducedby about 10% to about 30%. In certain embodiments, the fatigue isreduced by about 10% to about 25%. In certain embodiments, the fatigueis reduced by about 10% to about 20%. In certain embodiments, thefatigue is reduced by about 10% to about 15%. In certain of theforegoing embodiments, the subject has been treated with atrasentan or apharmaceutically acceptable salt thereof for between about 15 days andabout 30 days. In certain embodiments, the decrease in fatigue comprisesa decrease in the score on one or more of the Fatigue Severity Scale,the Chalder Fatigue Scale, the FACIT Fatigue Scale, the Brief FatigueInventory, the FACT-F Subscale, Global Vigor and Affect, the May andKline Adjective Checklist, the Pearson-Byars Fatigue Feeling Checklist,the Rhoten Fatigue Scale, the Schedule of Fatigue and Anergia, or theChecklist Individual Strength.

Some embodiments provide a method of inhibiting mesangial cellactivation in a subject having IgA nephropathy, comprising administeringa therapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, to the subject; wherein the subject has notbeen previously diagnosed with one or more of diabetic nephropathy,HIV/AIDS, or acute kidney failure.

Some embodiments provide a method of inhibiting PDGF signaling activity(e.g., decreasing the expression and/or activity of one or more ofPIK3R1, PDGFRA, NFKBIA, PIK3CG, PLA2G4A, TIAM1, PDGFB, NFKB1, andMAP3K1) in a mesangial cell in a subject having IgA nephropathy,comprising administering a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof, to thesubject; wherein the subject has not been previously diagnosed with oneor more of diabetic nephropathy, HIV/AIDS, or acute kidney failure.

Some embodiments provide a method of inhibiting mesangial cellactivation, comprising contacting a mesangial cell with an effectiveamount of atrasentan, or a pharmaceutically acceptable salt thereof.

In some embodiments, the mesangial activation is induced by IgA immunecomplexes. In some embodiments, the mesangial activation is associatedwith the presence of IgA immune complexes. The presence and/or amount ofIgA immune complexes can be detected by a variety of methods. Forexample, the complexes may be detected in serum or urine, and can alsobe detected in a kidney biopsy sample.

In some embodiments, the inhibiting of mesangial cell activationcomprises reducing expression and/or activity of one or more biomarkersindicative of mesangial cell proliferation. In some embodiments,inhibiting of mesangial cell activation comprises reducing mesangialcell inflammation. In some embodiments, reducing mesangial cellinflammation comprises reducing expression and/or activity of one ormore of IL6, MCP1, or other biomarkers indicative of mesangial cellinflammation. In some embodiments, reducing mesangial cell inflammationcomprises reducing expression and/or activity of IL-6. In someembodiments, the expression and/or activity of one or more biomarkersindicative of mesangial cell inflammation is reduced by about 25% toabout 99% after treatment with atrasentan or a pharmaceuticallyacceptable salt thereof (e.g., after treatment for about 1 week, about 2weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 20 weeks,about 30 weeks, about 40 weeks, about 50 weeks, about 60 weeks, about 70weeks, about 80 weeks, about 90 weeks, about 100 weeks, about 110 weeks,about 120 weeks, about 130 weeks, about 140 weeks, about 150 weeks,about 160 weeks, about 170 weeks, about 180 weeks, about 190 weeks, orabout 200 weeks, or any value in between). In some embodiments, theexpression and/or activity of one or more biomarkers indicative ofmesangial cell inflammation is reduced by about 25% to about 50%, about40% to about 60%, about 50% to about 75%, about 60% to about 80%, about75% to about 90%, about 85% to about 99%, or any value in between. Forexample, in some such embodiments, the one or more biomarkers can beIL-6.

In some embodiments, inhibiting of mesangial cell activation comprisesreducing mesangial cell inflammation. In some embodiments, reducingmesangial cell inflammation comprises reducing IL-6 signaling (e.g.,reducing the expression and/or activity in one or more proteins involvedin an IL-6 signaling pathway, e.g., a reduction in the expression and/oractivity of one or more of Cntfr, Il1b, Csf1, Il2ra, Map3k8, and Il1r1).In some embodiments, reducing mesanial cell inflammation comprisesreducing the expression and/or activity of one or more (e.g., 1, 2, 3,4, or 5) of: Cntfr, Il1b, Csf1, Il2ra, Map3k8, Il1r1.

In some embodiments, the inhibiting of mesangial cell activationcomprises reducing the pro-fibrotic response in the mesangial cells. Insome embodiments, reducing the pro-fibrotic response in the mesangialcells comprises reducing expression and/or activity of one or more ofNF-κB, TGF, PDGF, CTGF, MMP, TIMPS, or other biomarkers indicative ofmesangial cell fibrosis. In some embodiments, the expression and/oractivity of one or more of NF-κB, TGF, PDGF, CTGF, MMP, and TIMPS, isreduced by about 25% to about 99% after treatment with atrasentan or apharmaceutically acceptable salt thereof (e.g., after treatment forabout 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about10 weeks, about 20 weeks, about 30 weeks, about 40 weeks, about 50weeks, about 60 weeks, about 70 weeks, about 80 weeks, about 90 weeks,about 100 weeks, about 110 weeks, about 120 weeks, about 130 weeks,about 140 weeks, about 150 weeks, about 160 weeks, about 170 weeks,about 180 weeks, about 190 weeks, or about 200 weeks, or any value inbetween), relative to the expression and/or activity prior toadministration of atrasentan, or a pharmaceutically acceptable saltthereof. In some embodiments, the expression and/or activity of one ormore of NF-κB, TGF, PDGF, CTGF, MMP, and TIMPS is reduced by about 25%to about 50%, about 40% to about 60%, about 50% to about 75%, about 60%to about 80%, about 75% to about 90%, about 85% to about 99%, or anyvalue in between.

In some embodiments, inhibiting of mesangial cell activation comprisesreducing the pro-fibrotic response in the mesangial cells. In someembodiments, reducing the the pro-fibrotic response comprises reducingNF-κB signaling. In some embodiments, reducing the pro-fibrotic responsecomprises reducing the expression and/or activity of one or more (e.g.,1, 2, 3, 4, or 5) of: Pfkfb3, Nr4a1, Gem, Fosl2, Klf4, F3, Nfkbia,Ifit2, Nr4a2, Klf2, Jag1, Dnajb4, Il1b, Spsb1, Btg2, Atf3, Csf1, Trib1,Zbtb10, Btg1, Rhob, Nfat5, Edn1, Rel, Nr4a3, Nfkb1, Serpine1, Ccl20,Perl, Cxcl2, Map3k8, Traf1, and/or increasing the expression and/oractivity of one or more (e.g., 1, 2, 3, 4, or 5) of: Ehd1, Snn, Tnfaip8,Ackr3, Id2, Ccn1, Efna1, Ccnd1, Cdkn1a, Pnrc1 (in cases where thecomponent inhibits NF-κB signaling).

In some embodiments, reducing the pro-fibrotic response comprisesreducing PDGF signaling. In some embodiments, reducing the pro-fibroticresponse comprises reducing the expression and/or activity of one ormore (e.g., 1, 2, 3, 4, or 5) of: Pik3r1, Pdgfra, Nfkbia, Pik3cg,Pla2g4a, Tiam1, Pdgfb, Nfkb1, and/or increasing the expression and/oractivity of one or more (e.g., 1, 2, 3, 4, or 5) of: Hras (in caseswhere the component inhibits PDGF signaling).

In some embodiments, the expression and/or activity of NF-κB and/or PDGFexpression and/or activity, is reduced by about 25% to about 99% aftertreatment with atrasentan or a pharmaceutically acceptable salt thereof(e.g., after treatment for about 1 week, about 2 weeks, about 3 weeks,about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8weeks, about 9 weeks, about 10 weeks, about 20 weeks, about 30 weeks,about 40 weeks, about 50 weeks, about 60 weeks, about 70 weeks, about 80weeks, about 90 weeks, about 100 weeks, about 110 weeks, about 120weeks, about 130 weeks, about 140 weeks, about 150 weeks, about 160weeks, about 170 weeks, about 180 weeks, about 190 weeks, or about 200weeks, or any value in between), relative to the expression and/oractivity prior to administration of atrasentan, or a pharmaceuticallyacceptable salt thereof. In some embodiments, the expression and/oractivity of NF-κB and/or PDGF is reduced by about 25% to about 50%,about 40% to about 60%, about 50% to about 75%, about 60% to about 80%,about 75% to about 90%, about 85% to about 99%, or any value in between.

In some embodiments, reducing the pro-fibrotic response in the mesangialcells comprises reducing matrix secretion by mesangial cells. In someembodiments, reducing matrix secretion by mesangial cells comprisesreducing expression and/or activity of one or more of excess matrixsecretion by mesangial cells.

Some embodiments provide a method of reducing activation of a mesangialcell in contact with an IgA immune complex, comprising contacting amesangial cell with an effective amount of atrasentan, or apharmaceutically acceptable salt thereof. In some embodiments, reducingactivation of a mesangial cell comprises reducing expression and/oractivity of one or more biomarkers indicative of mesangial cellproliferation.

In some embodiments, reducing activation of a mesangial cell comprisesreducing mesangial cell inflammation. In some embodiments, reducingmesangial cell inflammation comprises reducing expression and/oractivity of one or more of IL6, MCP1, or other biomarkers indicative ofmesangial cell inflammation.

In some embodiments, reducing activation of a mesangial cell comprisesreducing the pro-fibrotic response in the mesangial cells. In someembodiments, reducing the pro-fibrotic response in the mesangial cellscomprises reducing expression and/or activity of one or more of TGF,PDGF, CTGF, MMP, TIMPS, or other biomarkers indicative of mesangial cellfibrosis.

In some embodiments, reducing the pro-fibrotic response in the mesangialcells comprises reducing matrix secretion by mesangial cells. In someembodiments, reducing matrix secretion by mesangial cells comprisesreducing expression and/or activity of one or more biomarkers indicativeof excess matrix secretion by mesangial cells.

In some embodiments, the reducing activation of a mesangial cellcomprises reducing undesired mesangial cell migration. In someembodiments, the reduction in undesired mesangial cell migration occursafter about 15 days to about 30 days after treatment with atrasentan, ora pharmaceutically acceptable salt thereof. In some embodiments, thereduction in undesired mesangial cell migration occurs after about 3months to about 6 months after treatment with atrasentan, or apharmaceutically acceptable salt thereof.

In some embodiments, the reducing activation of a mesangial cellcomprises reducing undesired mesangial cell proliferation. In someembodiments, the reduction in undesired mesangial cell proliferationoccurs after about 15 days to about 30 days after treatment withatrasentan, or a pharmaceutically acceptable salt thereof. In someembodiments, the reduction in undesired mesangial cell proliferationoccurs after about 3 months to about 6 months after treatment withatrasentan, or a pharmaceutically acceptable salt thereof.

In some embodiments, the undesired mesangial cell proliferation isreduced by about 25% to about 99% after treatment with atrasentan or apharmaceutically acceptable salt thereof (e.g., after treatment forabout 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about10 weeks, about 20 weeks, about 30 weeks, about 40 weeks, about 50weeks, about 60 weeks, about 70 weeks, about 80 weeks, about 90 weeks,about 100 weeks, about 110 weeks, about 120 weeks, about 130 weeks,about 140 weeks, about 150 weeks, about 160 weeks, about 170 weeks,about 180 weeks, about 190 weeks, or about 200 weeks, or any value inbetween). In some embodiments, the the undesired mesangial cellproliferation is reduced by about 25% to about 50%, about 40% to about60%, about 50% to about 75%, about 60% to about 80%, about 75% to about90%, about 85% to about 99%, or any value in between.

In some embodiments, mesangial cell activation can be assessed by one ormore of serum analysis, urinalysis, and microscopy of a kidney biopsysample (e.g., light microscopy and/or immunofluorescence microscopy).

In some embodiments, the contacting occurs in vitro. In someembodiments, the contacting occurs in vivo.

Some embodiments provide a method of treating IgA nephropathy in asubject in need thereof, comprising: a) determining that the subject haselevated serum Gd-IgA1 levels; and b) administering a therapeuticallyeffective amount of atrasentan, or a pharmaceutically acceptable saltthereof, to the subject. In some embodiments, the subject has not beenpreviously diagnosed with one or more of diabetic nephropathy, HIV/AIDS,or acute kidney failure. In some embodiments, the subject has not beenpreviously diagnosed with HIV-related nephropathy. In some embodiments,the subject has not been previously diagnosed with cancer. In someembodiments, the cancer is lung cancer or prostate cancer.

Some embodiments provide a method of treating IgA nephropathy in asubject in need thereof, comprising: a) determining that the subject haselevated levels of mesangial activation; and b) administering atherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, to the subject.

In some embodiments, determining of elevated levels of mesangialactivation comprises obtaining a sample from the subject and assessingthe level of mesangial activation in the same. In some embodiments, thesample is a kidney biopsy sample. In some embodiments, the sample isselected from a blood sample, a urine sample, a kidney biopsy sample, ora combination of two or three of the foregoing.

In some embodiments, the sample exhibits elevated levels of one or moreof: matrix secretion by the mesangial cells, IgA-immune complexdeposition, mesangial cell proliferation, and endocapillary cellproliferation. In some embodiments, the sample exhibits elevated levelsof IgA-immune complex deposition.

In some embodiments, the subject has been determined to have proteinuriaof at least about 1 g/day in at least two of three consecutive readingsover the year prior to administration of a therapeutically effectiveamount of atrasentan, or a pharmaceutically acceptable salt thereof. Forexample, about 1 g/day, about 1.2 g/day, about 1.4 g/day, about 1.6g/day, about 1.8 g/day, or about at least 2 g/day.

In some embodiments, the subject has been administered a maximallytolerated stable dose of a RAS inhibitor for at least 12 weeks prior toadministration of a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof. In some embodiments, thesubject is concurrently administered a maximally tolerated stable doseof a RAS inhibitor and a therapeutically effective amount of atrasentan,or a pharmaceutically acceptable salt thereof. In some embodiments, theRAS inhibitor is an angiotensin-converting enzyme inhibitor. In someembodiments, the RAS inhibitor is an angiotensin receptor blocker (ARB).

In some embodiments, the subject has been determined to have hematuriaprior to administration of a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof. In someembodiments, the hematuria is microhematuria. In some embodiments, thehematuria is gross hematuria.

In some embodiments, the subject has been determined to have an eGFR ofat least 30 mL/min/1.73 m² prior to administration of a therapeuticallyeffective amount of atrasentan, or a pharmaceutically acceptable saltthereof. In some embodiments, the subject has been determined to have aneGFR of about 30 mL/min/1.73 m² to about 60 mL/min/1.73 m² prior toadministration of a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof.

In some embodiments, the subject has not been previously diagnosed withone or more of diabetic nephropathy, HIV/AIDS, or acute kidney failure.In some embodiments, the subject has not been previously diagnosed withHIV-related nephropathy. In some embodiments, the subject has not beenpreviously diagnosed with cancer. In some embodiments, the cancer islung cancer or prostate cancer.

Some embodiments provide a method of treating IgA nephropathy in asubject in need thereof, comprising: a) determining that the subject haselevated levels of IgA-immune complexes in the kidney; and b)administering a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof, to the subject.

In some embodiments, determining of elevated levels of IgA-immunecomplexes in the kidney comprises obtaining a sample from the subjectand assessing the level of IgA-immune complexes in the same. In someembodiments, the sample is a kidney biopsy sample. In some embodiments,the sample is selected from a blood sample, a urine sample, a kidneybiopsy sample, or a combination of two or three of the foregoing. Insome embodiments, the IgA-immune complexes are deposited in themesangium.

In some embodiments, the levels of IgA-immune complexes can be assessedby one or more of serum analysis, urinalysis, and microscopy of a kidneybiopsy sample (e.g., light microscopy and/or immunofluorescencemicroscopy).

In some embodiments, the sample exhibits elevated levels of one or moreof: matrix secretion by the mesangial cells, IgA-immune complexdeposition in the mesangium, mesangial cell activation, mesangial cellproliferation, and endocapillary cell proliferation.

In some embodiments, the subject has been determined to have proteinuriaof at least about 1 g/day in at least two of three consecutive readingsover the year prior to administration of a therapeutically effectiveamount of atrasentan, or a pharmaceutically acceptable salt thereof. Forexample, about 1 g/day, about 1.2 g/day, about 1.4 g/day, about 1.6g/day, about 1.8 g/day, or about at least 2 g/day.

In some embodiments, the subject has been administered a maximallytolerated stable dose of a RAS inhibitor for at least 12 weeks prior toadministration of a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof. In some embodiments, thesubject is concurrently administered a maximally tolerated stable doseof a RAS inhibitor and a therapeutically effective amount of atrasentan,or a pharmaceutically acceptable salt thereof. In some embodiments, theRAS inhibitor is an angiotensin-converting enzyme inhibitor. In someembodiments, the RAS inhibitor is an angiotensin receptor blocker (ARB).

In some embodiments, the subject has been determined to have hematuriaprior to administration of a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof. In someembodiments, the hematuria is microhematuria. In some embodiments, thehematuria is gross hematuria.

In some embodiments, the subject has been determined to have an eGFR ofat least 30 mL/min/1.73 m² prior to administration of a therapeuticallyeffective amount of atrasentan, or a pharmaceutically acceptable saltthereof. In some embodiments, the subject has been determined to have aneGFR of about 30 mL/min/1.73 m² to about 60 mL/min/1.73 m² prior toadministration of a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof.

In some embodiments, the subject has not been previously diagnosed withone or more of diabetic nephropathy, HIV/AIDS, or acute kidney failure.In some embodiments, the subject has not been previously diagnosed withHIV-related nephropathy. In some embodiments, the subject has not beenpreviously diagnosed with cancer. In some embodiments, the cancer islung cancer or prostate cancer.

In some embodiments, the methods include determining, in the subject,expression and/or activity of one or more of ET1, TGF, PDGF, CTGF, MMP,TIMPS, IGF1, DPEP1, ASL, AMN, ALPL, SLC6A19, IL-6, and NF-kB. In someembodiments, expression and/or activity are determined prior toadministration of a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof. In some embodiments,expression and/or activity are determined after administration of atherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof.

In some embodiments, the determining the expression and/or activity isperformed prior to administration of a therapeutically effective amountof atrasentan, or a pharmaceutically acceptable salt thereof. In someembodiments, the determining the expression and/or activity is performedafter administration of a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof, for example,after treatment for about 1 week, about 2 weeks, about 3 weeks, about 4weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about9 weeks, about 10 weeks, about 20 weeks, about 30 weeks, about 40 weeks,about 50 weeks, about 60 weeks, about 70 weeks, about 80 weeks, about 90weeks, about 100 weeks, about 110 weeks, about 120 weeks, about 130weeks, about 140 weeks, about 150 weeks, about 160 weeks, about 170weeks, about 180 weeks, about 190 weeks, or about 200 weeks, or anyvalue in between.

In some embodiments, the subject has been determined to have elevatedexpression and/or activity of one or more of ET1, TGF, PDGF, CTGF, MMP,TIMPS, IGF1, DPEP1, ASL, AMN, ALPL, SLC6A19, IL-6, NF-kB, PKC, PI3K,Src, Ras, ERK1/2, Rho, Rac, Akt, mTOR, NAPDH oxidase, MAPK, cPLA2,TNF-α, IL-1, CAM, COX-2, iNOS, JAK, STAT3, PI3K, Akt/PKB, IKKs, IkBs,NF-kB, MAPK, Ras, Raf, MEK, ERK, MCP1, Cntfr, Il1b, Csf1, Il2ra, Map3k8,Il1r1, Pfkfb3, Nr4a1, Gem, Fosl2, Klf4, F3, Nfkbia, Ifit2, Nr4a2, Klf2,Jag1, Dnajb4, Il1b, Spsb1, Btg2, Atf3, Csf1, Trib1, Zbtb10, Btg1, Rhob,Nfat5, Edn1, Rel, Nr4a3, Nfkb1, Serpine1, Ccl20, Perl, Cxcl2, Map3k8,Traf1, Pik3r1, Pdgfra, Nfkbia, Pik3cg, Pla2g4a, Tiam1, and Pdgfb. Insome embodiments, the subject has been determined to have elevatedexpression and/or activity of one or more of ET1, TGF, PDGF, CTGF, MMP,TIMPS, IGF1, DPEP1, ASL, AMN, ALPL, SLC6A19, IL-6, NF-kB, PKC, PI3K,Src, Ras, ERK1/2, Rho, Rac, Akt, mTOR, NAPDH oxidase, MAPK, cPLA2,TNF-α, IL-1, CAM, COX-2, iNOS, JAK, STAT3, PI3K, Akt/PKB, IKKs, IkBs,NF-kB, MAPK, Ras, Raf, MEK, ERK, and MCP1. In some embodiments, thesubject has been determined to have elevated expression and/or activityof one or more of Cntfr, Il1b, Csf1, Il2ra, Map3k8, Il1r1, Pfkfb3,Nr4a1, Gem, Fosl2, Klf4, F3, Nfkbia, Ifit2, Nr4a2, Klf2, Jag1, Dnajb4,Il1b, Spsb1, Btg2, Atf3, Csf1, Trib1, Zbtb10, Btg1, Rhob, Nfat5, Edn1,Rel, Nr4a3, Nfkb1, Serpine1, Ccl20, Perl, Cxcl2, Map3k8, Traf1, Pik3r1,Pdgfra, Nfkbia, Pik3cg, Pla2g4a, Tiam1, and Pdgfb. In some embodiments,the subject has been determined to have elevated expression and/oractivity of one or more of ET1, TGF, PDGF, CTGF, MMP, TIMPS, IGF1,DPEP1, ASL, AMN, ALPL, SLC6A19, IL-6, and NF-kB. In some embodiments,the subject has been determined to have elevated expression and/oractivity of one or more of ET1, TGF, PDGF, CTGF, MMP, TIMPS, IGF1,DPEP1, ASL, AMN, ALPL, and SLC6A19.

Some embodiments provide a method of treating IgA nephropathy in asubject, comprising: (a) determining that the subject has elevatedexpression and/or activity of one or more of ET1, TGF, PDGF, CTGF, MMP,TIMPS, IGF1, DPEP1, ASL, AMN, ALPL, SLC6A19, IL-6, NF-kB, PKC, PI3K,Src, Ras, ERK1/2, Rho, Rac, Akt, mTOR, NAPDH oxidase, MAPK, cPLA2,TNF-α, IL-1, CAM, COX-2, iNOS, JAK, STAT3, PI3K, Akt/PKB, IKKs, IkBs,NF-kB, MAPK, Ras, Raf, MEK, ERK, MCP1, Cntfr, Il1b, Csf1, Il2ra, Map3k8,Il1r1, Pfkfb3, Nr4a1, Gem, Fosl2, Klf4, F3, Nfkbia, Ifit2, Nr4a2, Klf2,Jag1, Dnajb4, Il1b, Spsb1, Btg2, Atf3, Csf1, Trib1, Zbtb10, Btg1, Rhob,Nfat5, Edn1, Rel, Nr4a3, Nfkb1, Serpine1, Ccl20, Perl, Cxcl2, Map3k8,Traf1, Pik3r1, Pdgfra, Nfkbia, Pik3cg, Pla2g4a, Tiam1, and Pdgfb; and(b) administering to the subject a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof.

Some embodiments provide a method of treating IgA nephropathy in asubject determined to have elevated expression and/or activity of one ormore of ET1, TGF, PDGF, CTGF, MMP, TIMPS, IGF1, DPEP1, ASL, AMN, ALPL,SLC6A19, IL-6, NF-kB, PKC, PI3K, Src, Ras, ERK1/2, Rho, Rac, Akt, mTOR,NAPDH oxidase, MAPK, cPLA2, TNF-α, IL-1, CAM, COX-2, iNOS, JAK, STAT3,PI3K, Akt/PKB, IKKs, IkBs, NF-kB, MAPK, Ras, Raf, MEK, ERK, MCP1, Cntfr,Il1b, Csf1, Il2ra, Map3k8, Il1r1, Pfkfb3, Nr4a1, Gem, Fosl2, Klf4, F3,Nfkbia, Ifit2, Nr4a2, Klf2, Jag1, Dnajb4, Il1b, Spsb1, Btg2, Atf3, Csf1,Trib1, Zbtb10, Btg1, Rhob, Nfat5, Edn1, Rel, Nr4a3, Nfkb1, Serpine1,Ccl20, Perl, Cxcl2, Map3k8, Traf1, Pik3r1, Pdgfra, Nfkbia, Pik3cg,Pla2g4a, Tiam1, and Pdgfb, comprising administering to the subject atherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof.

D. Atrasentan

Atrasentan, also known as(2R,3R,4S)-4-(1,3-benzodioxol-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylicacid, ABT-627, A-147627, or A-127722, is a small molecule of thefollowing chemical structure:

Atrasaentan and methods of preparation thereof are described in U.S.Pat. No. 7,208,517 and International Patent Application Publication No.WO 1997/030045 (see e.g., Example 501), each of which is incorporatedherein by reference in its entirety.

In some embodiments, atrasentan is administered as a free base. In someother embodiments, atrasentan is administered as a pharmaceuticallyacceptable salt as described anywhere herein.

Atrasentan is an ET_(A) inhibitor which is about 1,860 times moreselective for ET_(A) relative to ET_(B). As used herein “ETA” is theabbreviation for endothelin receptor A; and “ET_(B)” is the abbreviationof endothelin receptor B. See, e.g., Ann Rheum Dis., 66(11), pp.1467-1472 (2007); Eur. Resp. J., 37, pp. 475-476 (2011); Plos One, 9,e87548 (2014); J. Clin. Oncol., 10, 31(14), pp. 1740-7 (2013);Pharmacol. Rev., 68 (2) pp. 357-418 (2016); and Nephrol. Dial.Transplant., 29, pp. i69-i73 (2014).

Salts

In some embodiments, atrasentan is in the form of a pharmaceuticallyacceptable salt. The phrase “pharmaceutically acceptable salt” as usedherein, refers to pharmaceutically acceptable organic or inorganic saltsof a compound of the disclosure (e.g., atrasentan). Exemplary saltsinclude acid addition salts formed by the reaction between atrasentanand an acid (e.g., organic acid or inorganic acid). Non-limitingexamples include: sulfate, citrate, acetate, oxalate, chloride, bromide,iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate,lactate, salicylate, acid citrate, tartrate, oleate, tannate,pantothenate, bitartrate, ascorbate, succinate, maleate, mandelate(e.g., (S)-mandelate or (R)-mandelate), gentisinate, fumarate,gluconate, glucuronate, saccharate, formate, benzoate, glutamate,methanesulfonate “mesylate”, ethanesulfonate, benzenesulfonate, andp-toluenesulfonate, pamoate (i.e.,4,4′-methylene-bis-(2-hydroxy-3-naphthoate)) salts. Exemplary salts alsoinclude base addition salts formed by the reaction between atrasentanand a base. Non-limiting examples include: alkali metal (e.g., sodiumand potassium) salts, alkaline earth metal (e.g., magnesium) salts, andammonium salts. A pharmaceutically acceptable salt may involve theinclusion of another molecule such as an acetate ion, a succinate ion orother counter ion. The counter ion may be any organic or inorganicmoiety that stabilizes the charge on the parent compound. Furthermore, apharmaceutically acceptable salt may have more than one charged atom inits structure. Instances where multiple charged atoms are part of thepharmaceutically acceptable salt can have multiple counter ions. Hence,a pharmaceutically acceptable salt can have one or more charged atomsand/or one or more counter ion. When referring to atrasentan, the term“salt” or “salts” is understood to be a salt of atrasentan that can bepresent alone or in a mixture with free atrasentan.

In some embodiments, atrasentan is in the form of a hydrochloride salt.The hydrochloride salt of atrasentan, also known as atrasentanhydrochloride (CAS Number: 195733-43-8); atrasentan hydrogen chloride;atrasentan hydrochloride salt; atrasentan chloride salt; atrasentan HCl;atrasentan monohydrochloride;(2R,3R,4S)-4-(1,3-benzodioxol-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylic acid, monohydrochloride;3-pyrrolidinecarboxylic acid,4-(1,3-benzodioxol-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)-,hydrochloride (1:1), (2R, 3R, 4S)—; (2R, 3R,4S)-1-[(dibutylcarbamoyl)methyl]-2-(p-methoxyphenyl)-4-[3,4-(methylenedioxy)phenyl]-3-pyrrolidinecarboxylicacid, monohydrochloride; 3-pyrrolidinecarboxylic acid,4-(1,3-benzodioxol-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)-,monohydrochloride, [2R-(2α, 3β, 4α)]; ABT-627; A-147627.1;Abbott-147627.1, has the following structure:

wherein the molar ratio of atrasentan to chloride is 1:1. Atrasentanhydrochloride and methods of preparation thereof are further describedin U.S. Pat. No. 7,208,517 and International Patent ApplicationPublication No. WO 1997/030045 (see e.g., Example 501), each of which isincorporated herein by reference in its entirety.

In some embodiments, atrasentan is in the form of a mandelate salt. Incertain embodiments, atrasentan is in the form of a (S)-mandelate salt.In certain embodiments, atrasentan is in the form of a (R)-mandelatesalt. In certain embodiments, in the atrasentan mandelate salt,atrasentan and mandelate has a molar ratio of 1:1. In certainembodiments, in the atrasentan mandelate salt, atrasentan and mandelatehas a molar ratio of 2:1. Atrasentan mandelate salt and methods ofpreparation thereof are further described in U.S. Pat. Nos. 8,962,675and 9,637,476, each of which is incorporated herein by reference in itsentirety.

In some embodiments, atrasentan is in the form of a hemisulfate salt.Hemisulfate salt and methods of preparation thereof are furtherdescribed in U.S. Pat. Nos. 8,962,675 and 9,637,476, each of which isincorporated herein by reference in its entirety.

In some embodiments, the atrasentan or a pharmaceutically acceptablesalt thereof is in the form of an anhydrate. In certain embodiments, theatrasentan or a pharmaceutically acceptable salt thereof is in the formof a hydrate. In certain embodiments, the atrasentan or apharmaceutically acceptable salt thereof is in the form of a solvate.

Stereochemistry

Atrasentan possesses three asymmetric centers and can be produced asindividual stereoisomers (e.g., enantiomers or diastereomers) or asmixtures thereof as described in U.S. Pat. No. 7,208,517 andInternational Patent Application Publication No. WO 1997/030045. In someembodiments, atrasentan as described herein comprises the(2R,3R,4S)-stereoisomer, that is(2R,3R,4S)-4-(1,3-benzodioxol-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylicacid. In certain embodiments, atrasentan is the (2R,3R,4S)-stereoisomerthat is substantially free of the other stereoisomers (e.g., contains<10%, <5%, <2%, <1%, <0.5%, <0.1%, or <0.05% of other stereoisomers).

Polymorphs

Atrasentan or a pharmaceutically acceptable salt thereof, as describedherein, can be in one or more polymorphic forms. In some embodiments,atrasentan or a pharmaceutically acceptable salt thereof issubstantially amorphous (e.g., >75%, >80%, >85%, >90%, >95%, >98%, >99%,or >99.5% amorphous). In some embodiments, atrasentan or apharmaceutically acceptable salt thereof is substantially crystalline(e.g., >75%, >80%, >85%, >90%, >95%, >98%, >99%, or >99.5% crystalline).

In certain embodiments, the atrasentan or a pharmaceutically acceptablesalt thereof comprises Atrasentan Hydrochloride Crystalline Form 1. Incertain embodiments, the atrasentan or a pharmaceutically acceptablesalt thereof is substantially Atrasentan Hydrochloride Crystalline Form1 (e.g., >75%, >80%, >85%, >90%, >95%, >98%, >99%, or >99.5% Form 1).Atrasentan Hydrochloride Crystalline Form 1 and methods of making thesame are described in International Patent Application Publication No.WO 2006/034094, which is incorporated by reference herein in itsentirety.

In some embodiments, Atrasentan Hydrochloride Crystalline Form 1 ischaracterized, when measured at about 25° C. with Cu-Kα radiation, by anX-ray powder diffraction pattern with at least three peaks (e.g., 3, 4,5, 6, or 7) having respective 2θ values of about 8.3°, 9.7°, 10.0°,13.0°, 15.6°, 17.2° or 19.5°. In certain embodiments, AtrasentanHydrochloride Crystalline Form 1 is characterized, when measured atabout 25° C. with Cu-Kα radiation, by an X-ray powder diffractionpattern with at least three peaks having respective 2θ values of about8.3°, 9.7°, 10.0°, 13.0°, 15.6°, 17.2° or 19.5°, and essentially withoutpeaks having 2θ values below about 6.2° and/or between about 6.6° and8.0°.

In some embodiments, Atrasentan Hydrochloride Crystalline Form 1 ischaracterized in the orthorhombic crystal system and P2₁2₁2₁ spacegroup, when measured at about 25° C. with Cu-Kα radiation, by latticeparameters a, b and c of 17.663 Å±0.005 Å, 21.24 Å±0.01 Å and 8.005Å±0.002 Å, respectively.

In some embodiments, Atrasentan Hydrochloride Crystalline Form 1 hassubstantial crystalline purity. In some embodiments, AtrasentanHydrochloride Crystalline Form 1 has substantial chemical purity. Insome embodiments, Atrasentan Hydrochloride Crystalline Form 1 hassubstantial diastereomeric purity.

Representative characteristic peak positions in the X-ray powderdiffraction pattern of Atrasentan Hydrochloride Crystalline Form I,expressed as degrees relative to 2θ, are, when measured at about 25° C.with Cu-Ka radiation, about 8.3° ((020), 77.35%); 9.7° ((120), 76.37%);10.0° ((200), 14.53%); 13.2° ((220), 28.03%); 13.6° ((130), 16.71%);14.9° ((121), 38.93%); 15.8° ((310), 13.11%); 16.2° ((230), 18.09%);17.4° ((320), 15.87%); 17.5° ((131), 37.80%); 19.6° ((240), 28.77%);20.8° ((141), 46.26%); 23.3° ((112), 100.0%); 24.3° ((151), 52.6%);25.3° ((341), 13.08%); and 25.9° ((132), 33.98%). Each peak position isshown with its accompanying Miller index (hkl) values and its integratedintensity (peak height). It is meant to be understood that peak heightsmay vary and will be dependent on variables such as the temperature,size of crystal size or morphology, sample preparation, or sample heightin the analysis well of the Scintag×2 Diffraction Pattern System. It isalso meant to be understood that peak positions may vary when measuredwith different radiation sources. For example, Cu-Kα1, Mo-Kα, Co-Kα andFe-Kα radiation, having wavelengths of 1.54060 Å, 0.7107 Å, 1.7902 Å and1.9373 Å, respectively, may provide peak positions that differ fromthose measured with Cu-Kα radiation.

In certain embodiments, the atrasentan or a pharmaceutically acceptablesalt thereof comprises Atrasentan Hydrochloride Crystalline Form 2. Incertain embodiments, the atrasentan or a pharmaceutically acceptablesalt thereof is substantially Atrasentan Hydrochloride Crystalline Form2 (e.g., >75%, >80%, >85%, >90%, >95%, >98%, >99%, or >99.5% Form 2).Atrasentan Hydrochloride Crystalline Form 2 and methods of making thesame are described in International Patent Application Publication No.WO 2006/034084, which is incorporated by reference herein in itsentirety.

In certain embodiments, Atrasentan Hydrochloride Crystalline Form 2 ischaracterized, when measured at about 25° C. with Cu-Kα radiation, by anX-ray powder diffraction pattern with peaks having respective 2θ valuesof about 6.7° and 22.05° and at least one peak having a respective 2θvalue of about 8.4°, 15.6°, 18.0°, 18.5°, 19.8° or 20.6°.

In certain embodiments, Atrasentan Hydrochloride Crystalline Form 2 hassubstantial crystalline purity and is characterized, when measured atabout 25° C. with Cu-Kα radiation, by an X-ray powder diffractionpattern with peaks having respective 2θ values of about 6.7° and 22.05°and at least one peak having a respective 2θ value of about 8.4°, 15.6°,18.0°, 18.5°, 19.8° or 20.6°.

In certain embodiments, Atrasentan Hydrochloride Crystalline Form 2 hassubstantial crystalline purity and substantial chemical purity; and saidAtrasentan Hydrochloride Crystalline Form 2 is characterized, whenmeasured at about 25° C. with Cu-Kα radiation, by an X-ray powderdiffraction pattern with peaks having respective 2θ values of about 6.7°and 22.05° and at least one peak having a respective 2θ value of about8.4°, 15.6°, 18.0°, 18.5°, 19.8° or 20.6°.

In certain embodiments, Atrasentan Hydrochloride Crystalline Form 2 hassubstantial crystalline purity, substantial chemical purity, andsubstantial diastereomeric purity; and said Atrasentan HydrochlorideCrystalline Form 2 is characterized, when measured at about 25° C. withCu-Kα radiation, by an X-ray powder diffraction pattern with peakshaving respective 2θ values of about 6.7° and 22.05° and at least onepeak having a respective 2θ value of about 8.4°, 15.6°, 18.0°, 18.5°,19.8° or 20.6°.

In certain embodiments, the atrasentan or a pharmaceutically acceptablesalt thereof comprises Atrasentan Hydrochloride Crystalline Form 3. Incertain embodiments, the atrasentan or a pharmaceutically acceptablesalt thereof is substantially Atrasentan Hydrochloride Crystalline Form3 (e.g., >75%, >80%, >85%, >90%, >95%, >98%, >99%, or >99.5% Form 3).Atrasentan Hydrochloride Crystalline Form 3 and methods of making thesame are described in International Patent Application Publication No.WO 2006/034234 and U.S. Pat. No. 9,051,301, each of which isincorporated by reference herein in its entirety.

In certain embodiments, Atrasentan Hydrochloride Crystalline Form 3 ischaracterized, when measured at about 25° C. with Cu-Kα radiation, by anX-ray powder diffraction pattern with peaks having respective 2θ valuesof about 6.7° and 21.95° and at least one peak having a respective 2θvalue of about 8.4°, 15.6°, 18.0°, 18.5°, 19.8° or 20.6°.

In certain embodiments, Atrasentan Hydrochloride Crystalline Form 3 hassubstantial crystalline purity and is characterized, when measured atabout 25° C. with Cu-Kα radiation, by an X-ray powder diffractionpattern with peaks having respective 2θ values of about 6.7° and 21.95°and at least one peak having a respective 2θ value of about 8.4°, 15.6°,18.0°, 18.5°, 19.8° or 20.6°.

In certain embodiments, Atrasentan Hydrochloride Crystalline Form 3 hassubstantial crystalline purity and substantial chemical purity; and saidAtrasentan Hydrochloride Crystalline Form 3 is characterized, whenmeasured at about 25° C. with Cu-Kα radiation, by an X-ray powderdiffraction pattern with peaks having respective 2θ values of about 6.7°and 21.95° and at least one peak having a respective 2θ value of about8.4°, 15.6°, 18.0°, 18.5°, 19.8° or 20.6°.

In certain embodiments, Atrasentan Hydrochloride Crystalline Form 3 hassubstantial crystalline purity, substantial chemical purity, andsubstantial diastereomeric purity; and said Atrasentan HydrochlorideCrystalline Form 3 is characterized, when measured at about 25° C. withCu-Kα radiation, by an X-ray powder diffraction pattern with peakshaving respective 2θ values of about 6.7° and 21.95° and at least onepeak having a respective 2θ value of about 8.4°, 15.6°, 18.0°, 18.5°,19.8° or 20.6°.

In certain embodiments, the atrasentan or a pharmaceutically acceptablesalt thereof comprises amorphous atrasentan hydrochloride. In certainembodiments, atrasentan hydrochloride is substantially amorphous(e.g., >75%, >80%, >85%, >90%, >95%, >98%, >99%, or >99.5% amorphous).Amorphous atrasentan hydrochloride and methods of making the same aredescribed in International Patent Application Publication No. WO2006/034085, which is incorporated by reference herein in its entirety.

In certain embodiments, the amorphous atrasentan hydrochloride hassubstantial chemical purity. In certain embodiments, the amorphousatrasentan hydrochloride has substantial diastereomeric purity.

In some embodiments, the atrasentan or a pharmaceutically acceptablesalt thereof comprises a crystalline atrasentan mandelate salt. Incertain embodiments, the atrasentan or a pharmaceutically acceptablesalt thereof is substantially a crystalline atrasentan mandelate salt(e.g., >75%, >80%, >85%, >90%, >95%, >98%, >99%, or >99.5% crystallineatrasentan mandelate salt).

In certain embodiments, the crystalline atrasentan mandelate salt is acrystalline atrasentan (S)-mandelate salt. In certain embodiments, theatrasentan (S)-mandelate salt is an anhydrous salt. In certainembodiments, the atrasentan (S)-mandelate salt is a solvated salt. Incertain embodiments, the atrasentan (S)-mandelate salt is a solvatedsalt selected from the group consisting of an acetonitrile solvate, anethanol solvate, and a pyridine solvate. In certain embodiments, theatrasentan (S)-mandelate salt is a hydrated salt.

(a) (S)-Mandelate Salt (1:1 Stoichiometry)

In certain embodiments, the crystalline atrasentan (S)-mandelate salt isa crystalline atrasentan (S)-mandelate salt wherein the molar ratio ofatrasentan to (S)-mandelate is about 1:1. In certain embodiments, theatrasentan (S)-mandelate salt is an anhydrous salt. In certainembodiments, the atrasentan (S)-mandelate salt is a solvated salt. Incertain embodiments, the atrasentan (S)-mandelate salt is a solvatedsalt selected from the group consisting of an acetonitrile solvate, anethanol solvate, and a pyridine solvate. In certain embodiments, theatrasentan (S)-mandelate salt is a hydrated salt. In certainembodiments, the atrasentan or a pharmaceutically acceptable saltthereof is substantially(e.g., >75%, >80%, >85%, >90%, >95%, >98%, >99%, or >99.5%) acrystalline atrasentan (S)-mandelate salt wherein the molar ratio ofatrasentan to (S)-mandelate is about 1:1.

In certain embodiments, the crystalline (S)-mandelate salt has an X-raypowder diffraction pattern comprising peaks at 5.5±0.2, 9.7±0.2, and19.4±0.2 degrees 2θ when measured at about 25° C. with monochromatic Kα1radiation. In certain embodiments, the crystalline (S)-mandelate salthas an X-ray powder diffraction pattern comprising peaks at 5.5±0.2,9.7±0.2, 12.1±0.2, and 19.4±0.2 degrees 2θ when measured at about 25° C.with monochromatic Kα1 radiation. In certain embodiments, thecrystalline (S)-mandelate salt has an X-ray powder diffraction patterncomprising peaks at 5.5±0.2, 9.7±0.2, 12.1±0.2, 18.0±0.2, 18.4±0.2, and19.4±0.2 degrees 2θ when measured at about 25° C. with monochromatic Kα1radiation. In certain embodiments, the experimental error associatedwith the X-ray powder diffraction peak values recited in the variousembodiments above is ±0.1 degrees 2θ. In certain embodiments, thecrystalline (S)-mandelate salt is an anhydrous salt. In certainembodiments, the molar ratio of atrasentan to (S)-mandelate is about1:1.

In certain embodiments, the crystalline (S)-mandelate salt has anorthorhombic lattice type. In certain embodiments, the crystalline(S)-mandelate salt has a P212121 space group. In certain embodiments,the crystalline (S)-mandelate salt has unit cell a, b and c values ofabout 9.954 Å, about 11.049 Å, and about 30.861 Å, respectively. Incertain embodiments, the crystalline (S)-mandelate salt has unit cell α,β and γ values of about 90°, about 90°, and about 90°, respectively. Incertain embodiments, the crystalline (S)-mandelate salt has at leastthree or more of the following properties: (a) an orthorhombic latticetype, (b) a P212121 space group, (c) unit cell a, b and c values ofabout 9.954 Å, about 11.049 Å, and about 30.861 Å, respectively, and/or(d) unit cell α, β and γ values of about 90°, about 90°, and about 90°,respectively. In certain embodiments, the crystalline (S)-mandelate salthas: (a) an orthorhombic lattice type, (b) a P212121 space group, (c)unit cell a, b and c values of about 9.954 Å, about 11.049 Å, and about30.861 Å, respectively, and (d) unit cell α, β and γ values of about90°, about 90°, and about 90°, respectively. In certain embodiments, thecrystalline (S)-mandelate salt is an anhydrous salt. In certainembodiments, the molar ratio of atrasentan to (S)-mandelate is about1:1.

(b) (S)-Mandelate Salt (2:1 Stoichiometry)

In certain embodiments, the crystalline (S)-mandelate salt is acrystalline atrasentan (S)-mandelate salt wherein the molar ratio ofatrasentan to (S)-mandelate is about 2:1. In certain embodiments, thecrystalline atrasentan (S)-mandelate salt is an anhydrous salt. Incertain embodiments, the crystalline atrasentan (S)-mandelate salt is asolvated salt. In certain embodiments, the crystalline atrasentan(S)-mandelate salt is a hydrated salt. In certain embodiments, theatrasentan or a pharmaceutically acceptable salt thereof issubstantially (e.g., >75%, >80%, >85%, >90%, >95%, >98%, >99%,or >99.5%) a crystalline atrasentan (S)-mandelate salt wherein the molarratio of atrasentan to (S)-mandelate is about 2:1.

In certain embodiments, the crystalline (S)-mandelate salt has an X-raypowder diffraction pattern comprising peaks at 4.5±0.2, 8.6±0.2, and18.1±0.2 degrees 2θ when measured at about 25° C. with monochromatic Kα1radiation. In certain embodiments, the crystalline (S)-mandelate salthas an X-ray powder diffraction pattern comprising peaks at 4.5±0.2,8.6±0.2, 18.1±0.2, and 18.7±0.2 degrees 2θ when measured at about 25° C.with monochromatic Kα1 radiation. In certain embodiments, thecrystalline (S)-mandelate salt has an X-ray powder diffraction patterncomprising peaks at 4.5±0.2, 8.6±0.2, 9.1±0.2, 18.1±0.2, and 18.7±0.2degrees 2θ when measured at about 25° C. with monochromatic Kα1radiation. In certain embodiments, the experimental error associatedwith the X-ray powder diffraction peak values recited in the variousembodiments above is ±0.1 degrees 2θ. In certain embodiments, thecrystalline (S)-mandelate salt is an anhydrous salt. In certainembodiments, the crystalline (S)-mandelate salt is a hydrated salt.

In certain embodiments, the crystalline atrasentan mandelate salt is acrystalline atrasentan (R)-mandelate salt. In certain embodiments, thecrystalline atrasentan (R)-mandelate salt is an anhydrous salt. Incertain embodiments, the crystalline atrasentan (R)-mandelate salt is asolvated salt. In certain embodiments, the crystalline atrasentan(R)-mandelate salt is a hydrated salt.

-   -   (c) (R)-Mandelate Salt (1:1 Stoichiometry)

In certain embodiments, the crystalline atrasentan (R)-mandelate salt isa crystalline atrasentan (R)-mandelate salt wherein the molar ratio ofatrasentan to (R)-mandelate is about 1:1. In certain embodiments, thecrystalline atrasentan (R)-mandelate salt is an anhydrous salt. Incertain embodiments, the crystalline atrasentan (R)-mandelate salt is asolvated salt. In certain embodiments, the crystalline atrasentan(R)-mandelate salt is a hydrated salt. In certain embodiments, theatrasentan or a pharmaceutically acceptable salt thereof issubstantially (e.g., >75%, >80%, >85%, >90%, >95%, >98%, >99%,or >99.5%) a crystalline atrasentan (R)-mandelate salt wherein the molarratio of atrasentan to (R)-mandelate is about 1:1.

In certain embodiments, the crystalline atrasentan (R)-mandelate salthas an X-ray powder diffraction pattern comprising peaks at 5.7±0.2,11.8±0.2, and 20.9±0.2 degrees 2θ when measured at about 25° C. withmonochromatic Kα1 radiation. In certain embodiments, the crystallineatrasentan (R)-mandelate salt has an X-ray powder diffraction patterncomprising peaks at 5.7±0.2, 8.2±0.2, 11.8±0.2, and 20.9±0.2 degrees 2θwhen measured at about 25° C. with monochromatic Kα1 radiation. Incertain embodiments, the crystalline atrasentan (R)-mandelate salt hasan X-ray powder diffraction pattern comprising peaks at 5.7±0.2,8.2±0.2, 8.6±0.2, 11.8±0.2, and 20.9±0.2 degrees 2θ when measured atabout 25° C. with monochromatic Kα1 radiation. In certain embodiments,the experimental error associated with the X-ray powder diffraction peakvalues recited in the various embodiments above is ±0.1 degrees 2θ. Incertain embodiments, the crystalline atrasentan (R)-mandelate salt is ananhydrous salt.

In some embodiments, the atrasentan or a pharmaceutically acceptablesalt thereof comprises an amorphous atrasentan mandelate salt. Incertain embodiments, the atrasentan or a pharmaceutically acceptablesalt thereof is substantially an amorphous atrasentan mandelate salt(e.g., >75%, >80%, >85%, >90%, >95%, >98%, >99%, or >99.5% amorphousatrasentan mandelate salt).

In certain embodiments, the amorphous atrasentan mandelate salt isamorphous atrasentan (S)-mandelate salt. In certain embodiments, theamorphous atrasentan (S)-mandelate salt is an anhydrous salt. In certainembodiments, the amorphous atrasentan (S)-mandelate salt is a solvatedsalt. In certain embodiments, the amorphous atrasentan (S)-mandelatesalt is a solvated salt selected from the group consisting of anacetonitrile solvate, an ethanol solvate, and a pyridine solvate. Incertain embodiments, the amorphous atrasentan (S)-mandelate salt is ahydrated salt. In certain embodiments, in the amorphous atrasentan(S)-mandelate salt, the molar ratio of atrasentan and (S)-mandelate isabout 1:1. In certain embodiments, in the amorphous atrasentan(S)-mandelate salt, the molar ratio of atrasentan and (S)-mandelate isabout 2:1.

In certain embodiments, the amorphous atrasentan mandelate salt isamorphous atrasentan (R)-mandelate salt. In certain embodiments, theamorphous atrasentan (R)-mandelate salt is an anhydrous salt. In certainembodiments, the amorphous atrasentan (R)-mandelate salt is a solvatedsalt. In certain embodiments, the amorphous atrasentan (R)-mandelatesalt is a solvated salt selected from the group consisting of anacetonitrile solvate, an ethanol solvate, and a pyridine solvate. Incertain embodiments, the amorphous atrasentan (R)-mandelate salt is ahydrated salt. In certain embodiments, in the amorphous atrasentan(R)-mandelate salt, the molar ratio of atrasentan and (R)-mandelate isabout 1:1. In certain embodiments, in the amorphous atrasentan(R)-mandelate salt, the molar ratio of atrasentan and (R)-mandelate isabout 2:1.

Crystalline and amorphous atrasentan mandelate salts are furtherdescribed in U.S. Pat. Nos. 8,962,675 and 9,637,476, each of which isincorporated herein by reference in its entirety.

E. Formulation

The term “pharmaceutical composition” as used herein is intended toencompass a product comprising the active ingredient(s), and the inertingredient(s) that make up the carrier, as well as any product whichresults, directly or indirectly, from combination, complexation oraggregation of any two or more of the ingredients, or from dissociationof one or more of the ingredients, or from other types of reactions orinteractions of one or more of the ingredients. Accordingly, thepharmaceutical compositions of the present disclosure encompass anycomposition made by admixing a compound of the present disclosure, or apharmaceutically acceptable salt, or solvate or solvate of the saltthereof, and a pharmaceutically acceptable carrier.

The amount administered depends on the compound formulation, route ofadministration, etc. and is generally empirically determined, andvariations will necessarily occur depending on the target, the host, andthe route of administration, etc. Generally, the quantity of activecompound in a unit dose of preparation may be varied or adjusted fromabout 0.1 milligram (mg) to about 10 mg or from about 0.5 mg to about 2mg, according to the particular application. For convenience, the totaldaily dosage may be divided and administered in portions during the day.

Pharmaceutical compositions of the present disclosure for injectioncomprise pharmaceutically acceptable sterile aqueous or non-aqueoussolutions, dispersions, suspensions or emulsions as well as sterilepowders for reconstitution into sterile injectable solutions ordispersions just prior to use. Examples of suitable aqueous andnon-aqueous carriers, diluents, solvents or vehicles include water,ethanol, polyols (such as glycerol, propylene glycol, polyethyleneglycol, and the like), and suitable mixtures thereof, vegetable oils(such as olive oil), and injectable organic esters such as ethyl oleate.Proper fluidity can be maintained, for example, by the use of coatingmaterials such as lecithin, by the maintenance of the required particlesize in the case of dispersions, and by the use of surfactants.

These pharmaceutical compositions may also contain adjuvants such aspreservative, wetting agents, emulsifying agents, and dispersing agents.Prevention of the action of micro-organisms may be ensured by theinclusion of various antibacterial and antifungal agents, for example,paraben, chlorobutanol, phenol sorbic acid, and the like. It may also bedesirable to include isotonic agents such as sugars, sodium chloride,and the like. Prolonged absorption of the injectable pharmaceutical formmay be brought about by the inclusion of agents that delay absorptionsuch as aluminum monostearate and gelatin. The compounds can beincorporated into slow release or targeted delivery systems such aspolymer matrices, liposomes, and microspheres. Such formulations mayprovide more effective distribution of the compounds.

The pharmaceutical compositions that are injectable formulations can besterilized, for example, by filtration through a bacterial-retainingfilter, or by incorporating sterilizing agents in the form of sterilesolid pharmaceutical compositions that can be dissolved or dispersed insterile water or other sterile injectable medium prior to use.

Solid dosage forms of the instant pharmaceutical compositions for oraladministration include capsules, tablets, pills, powders, and granules.In such solid dosage forms, the active compound is mixed with at leastone inert, pharmaceutically acceptable excipient or carrier such assodium citrate or dicalcium phosphate and/or a) fillers or extenderssuch as starches, lactose, sucrose, glucose, mannitol, and silicic acid,b) binders such as, for example, carboxymethylcellulose, alginates,gelatin, polyvinylpyrrolidone, sucrose, and acacia, c) humectants suchas glycerol, d) disintegrating agents such as agar-agar, calciumcarbonate, potato or tapioca starch, alginic acid, certain silicates,and sodium carbonate, e) solution retarding agents such as paraffin, f)absorption accelerators such as quaternary ammonium compounds, g)wetting agents such as, for example, cetyl alcohol and glycerolmonostearate, h) absorbents such as kaolin and bentonite clay, and i)lubricants such as talc, calcium stearate, magnesium stearate, solidpolyethylene glycols, sodium lauryl sulfate, and mixtures thereof. Inthe case of capsules, tablets and pills, the dosage form may alsocomprise buffering agents.

Solid pharmaceutical compositions of a similar type may also be employedas fillers in soft and hard-filled gelatin capsules using suchexcipients as lactose or milk sugar as well as high molecular weightpolyethylene glycols and the like.

The solid dosage forms of the instant pharmaceutical compositions oftablets, dragées, capsules, pills, and granules can be prepared withcoatings and shells such as enteric coatings and other pharmaceuticalcoatings. They may optionally contain opacifying agents and can also beof a formulation that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding pharmaceutical compositionswhich can be used include polymeric substances and waxes.

The active compounds can also be in microencapsulated form, ifappropriate, with one or more of the above-mentioned excipients.

Liquid dosage forms of the instant pharmaceutical compositions for oraladministration include pharmaceutically acceptable emulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethyl formamide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof.

Besides inert diluents, the oral pharmaceutical compositions can alsoinclude adjuvants such as wetting agents, emulsifying and suspendingagents, sweetening, flavoring, and perfuming agents.

Suspensions of the instant compounds, in addition to the activecompounds, may contain suspending agents as, for example, ethoxylatedisostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters,microcrystalline cellulose, aluminum metahydroxide, bentonite,agar-agar, and tragacanth, and mixtures thereof.

The compounds and compositions described herein can, for example, beadministered orally or parenterally, with a dosage ranging from about0.01 milligrams per kilogram (mg/kg) to about 0.05 mg/kg, every 4 to 120hours, or according to the requirements of the particular drug, dosageform, and/or route of administration. The interrelationship of dosagesfor animals and humans (based on milligrams per meter squared of bodysurface) is described by Freireich et al., Cancer Chemother. Rep. 50,219-244 (1966). Body surface area may be approximately determined fromheight and weight of the patient. See, e.g., Scientific Tables, GeigyPharmaceuticals, Ardsley, N.Y., 537 (1970). In certain embodiments, thecompositions are administered by oral administration or by injection.The methods herein contemplate administration of an effective amount ofcompound or compound composition to achieve a desired or stated effect.Typically, the pharmaceutical compositions of the present disclosurewill be administered from about 1 to about 6 times per day oralternatively, as a continuous infusion. Such administration can be usedas a chronic or acute therapy.

Lower or higher doses than those recited above may be required. Specificdosage and treatment regimens for any particular patient will dependupon a variety of factors, including the activity of the specificcompound employed, the age, body weight, general health status, sex,diet, time of administration, rate of excretion, drug combination, theseverity and course of the disease, condition or symptoms, the patient'sdisposition to the disease, and the judgment of the treating physician.

Dosage forms include from about 0.01 mg to about 10 mg (including, fromabout 0.1 mg to about 5 mg, from about 0.2 mg to about 4 mg, from about0.3 mg to about 3 mg, from about 0.4 mg to about 2 mg, from about 0.5 mgto about 1.5 mg, or from about 0.6 mg to about 1 mg) of a compound ofatrasentan, or a pharmaceutically acceptable salt thereof. In someembodiments, the dosage form include about 0.1 mg, about 0.2 mg, about0.3 mg, about 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.65, about 0.7mg, about 0.75, about 0.8 mg, about 0.85, about 0.9 mg, about 1 mg,about 1.1 mg, about 1.2 mg, about 1.3 mg, about 1.4 mg, about 1.5 mg,about 1.6 mg, about 1.7 mg, about 1.8 mg, about 1.9 mg, about 2 mg, orany value in between, of atrasentan, or a pharmaceutically acceptablesalt thereof. In some embodiments, the dosage form includes about 0.75mg of a compound of atrasentan, or a pharmaceutically acceptable saltthereof.

The dosage forms can further include a pharmaceutically acceptablecarrier and/or an additional therapeutic agent.

Appropriate dosage levels may be determined by any suitable method.Preferably, the active substance is administered at a frequency of 1 to4 times per day for topical administration, or less often if a drugdelivery system is used. Nevertheless, actual dosage levels and timecourse of administration of the active ingredients in the pharmaceuticalcompositions of the present disclosure may be varied so as to obtain anamount of the active ingredient which is effective to achieve a desiredtherapeutic response for a particular patient, composition and mode ofadministration, without being intolerably toxic to the patient. Incertain cases, dosages may deviate from the stated amounts, inparticular as a function of age, gender, body weight, diet and generalhealth status of the patient, route of administration, individualresponse to the active ingredient, nature of the preparation, and timeor interval over which administration takes place. Thus, it may besatisfactory in some cases to manage with less than the aforementionedminimum amount, whereas in other cases the stated upper limit may beexceeded. It may in the event of administration of larger amounts beadvisable to divide these into multiple individual doses spread over theday.

Exemplary Dosage Forms of Atrasentan

In some embodiments, provided herein a stable solid pharmaceuticaldosage form comprising: (a) about 0.25 mg to about 1.25 mg ofatrasentan, or an equivalent amount of a pharmaceutically acceptablesalt thereof; wherein the weight percent of atrasentan, orpharmaceutically acceptable salt thereof, in the dosage form is fromabout 0.05 weight percent to about 2.0 weight percent on an atrasentanfree base equivalent weight basis; and (b) a pharmaceutically acceptablediluent.

In some embodiments, provided herein a stable solid pharmaceuticaldosage form comprising: (a) about 0.25 mg to about 1.25 mg ofatrasentan, or an equivalent amount of a pharmaceutically acceptablesalt thereof; wherein the weight percent of atrasentan, orpharmaceutically acceptable salt thereof, in the dosage form is fromabout 0.05 weight percent to about 2.0 weight percent on an atrasentanfree base equivalent weight basis; (b) a pharmaceutically acceptableanti-oxidant; wherein the molar ratio of the anti-oxidant to atrasentan,or pharmaceutically acceptable salt thereof, is from about 10:1 to about1:10; and (c) a pharmaceutically acceptable diluent.

In certain of these embodiments, degradation of atrasentan in the dosageform is less than degradation of atrasentan in an otherwise identicaldosage form lacking the anti-oxidant when the dosage forms are storedfor a storage period of six months at about 40° C. and about 75%relative humidity.

In some embodiments, the dosage form is stored during the storage periodin a semi-permeable container or a substantially impermeable container.In some embodiments, the dosage form is stored during the storage periodin a sealed HDPE bottle or a blister package. In some embodiments, thedosage form is stored during the storage period in a sealed HDPE bottle.In some embodiments, the dosage form is stored during the storage periodin a blister package.

(i) Atrasentan

The dosage form can comprise a free base of atrasentan, apharmaceutically acceptable salt of atrasentan, or a combinationthereof. In some embodiments, the dosage form comprises a free base ofatrasentan. In some embodiments, the dosage form comprises apharmaceutically acceptable salt of atrasentan. In some embodiments, thedosage form comprises atrasentan hydrochloride. In some embodiments, thedosage form comprises atrasentan hydrochloride having a polymorph formselected from the group consisting of amorphous atrasentanhydrochloride, Atrasentan Hydrochloride Crystalline Form 1, AtrasentanHydrochloride Crystalline Form 2, and Atrasentan HydrochlorideCrystalline Form 3. In some embodiments, the dosage form comprisesamorphous atrasentan hydrochloride. In some embodiments, the dosage formcomprises Atrasentan Hydrochloride Crystalline Form 1. In someembodiments, the dosage form comprises Atrasentan HydrochlorideCrystalline form 2. In some embodiments, the dosage form comprisesatrasentan hydrochloride crystalline form 3. In some embodiments, thedosage form comprises atrasentan mandelate. In certain embodiments, thedosage form comprises a crystalline atrasentan mandelate (e.g., acrystalline atrasentan (S)-mandelate and/or a crystalline atrasentan(R)-mandelate). In certain embodiments, the dosage form comprises anamorphous atrasentan mandelate (e.g., an amorphous atrasentan(S)-mandelate and/or an amorphous atrasentan (R)-mandelate). In certainof the foregoing embodiments (when the dosage form comprises acrystalline and/or amorphous atrasentan (S)- and/or (R)-mandelate), themolar ratio of atrasentan and mandelate is 1:1. In certain otherembodiments, the molar ratio of atrasentan and mandelate is 2:1.

In certain embodiments, the dosage form comprises amorphous atrasentanhydrochloride; and it is substantially free (e.g., contains <10%, <5%,<1%, <0.5%, <0.1%, <0.05%) of other forms (e.g., other salts and/orother polymorphs) of atrasentan. In certain embodiments, the dosage formcomprises Atrasentan Hydrochloride Crystalline Form 1; and it issubstantially free of (e.g., contains <10%, <5%, <1%, <0.5%, <0.1%,<0.05%) other forms (e.g., other salts and/or other polymorphs) ofatrasentan. In certain embodiments, the dosage form comprises AtrasentanHydrochloride Crystalline form 2; and it is substantially free of otherforms (e.g., other salts and/or other polymorphs) of atrasentan. Incertain embodiments, the dosage form comprises atrasentan hydrochloridecrystalline form 3; and it is substantially free (e.g., contains <10%,<5%, <1%, <0.5%, <0.1%, <0.05%) of other forms (e.g., other salts and/orother polymorphs) of atrasentan. In certain embodiments, the dosage formcomprises crystalline atrasentan (S)-mandelate; and it is substantiallyfree (e.g., contains <10%, <5%, <1%, <0.5%, <0.1%, <0.05%) of otherforms (e.g., other salts and/or other polymorphs) of atrasentan. Incertain embodiments, the dosage form comprises crystalline atrasentan(R)-mandelate; and it is substantially free (e.g., contains <10%, <5%,<1%, <0.5%, <0.1%, <0.05%) of other forms (e.g., other salts and/orother polymorphs) of atrasentan. In certain embodiments, the dosage formcomprises amorphous atrasentan (S)-mandelate; and it is substantiallyfree (e.g., contains <10%, <5%, <1%, <0.5%, <0.1%, <0.05%) of otherforms (e.g., other salts and/or other polymorphs) of atrasentan. Incertain embodiments, the dosage form comprises amorphous atrasentan(R)-mandelate; and it is substantially free (e.g., contains <10%, <5%,<1%, <0.5%, <0.1%, <0.05%) of other forms (e.g., other salts and/orother polymorphs) of atrasentan.

In some embodiments, the weight percent of atrasentan, orpharmaceutically acceptable salt thereof, in the dosage form is fromabout 0.1 weight percent to about 2.0 weight percent on an atrasentanfree base equivalent weight basis. In some embodiments, the weightpercent of atrasentan, or pharmaceutically acceptable salt thereof, inthe dosage form is from about 0.2 weight percent to about 1.0 weightpercent on an atrasentan free base equivalent weight basis. In someembodiments, the weight percent of atrasentan, or pharmaceuticallyacceptable salt thereof, in the dosage form is from about 0.3 weightpercent to about 0.8 weight percent on an atrasentan free baseequivalent weight basis. In some embodiments, the weight percent ofatrasentan, or pharmaceutically acceptable salt thereof, in the dosageform is from about 0.40 weight percent to about 0.45 weight percent onan atrasentan free base equivalent weight basis. In some embodiments,the weight percent of atrasentan, or pharmaceutically acceptable saltthereof, in the dosage form is from about 0.60 weight percent to about0.65 weight percent on an atrasentan free base equivalent weight basis.

In some embodiments, the dosage form comprises from about 0.40 mg toabout 1.00 mg of atrasentan, or an equivalent amount of apharmaceutically acceptable salt thereof. In some embodiments, thedosage form comprises from about 0.40 mg to about 0.85 mg of atrasentan,or an equivalent amount of a pharmaceutically acceptable salt thereof.In some embodiments, the dosage form comprises about 0.50 mg ofatrasentan, or an equivalent amount of a pharmaceutically acceptablesalt thereof. In some embodiments, the dosage form comprises about 0.75mg of atrasentan, or an equivalent amount of a pharmaceuticallyacceptable salt thereof.

(ii) Diluent

Suitable diluents for use in the disclosed dosage forms include, but arenot limited to, lactose (such as lactose monohydrate, lactose anhydrous,and PHARMATOSE® DCL21), sucrose, glucose, mannitol, sorbitol, isomalt,microcrystalline cellulose (such as AVICEL® PH101 and AVICEL® PH102),silicified microcrystalline cellulose (such as PROSOLV® SMCC 50 and SMCC90), dicalcium phosphate, starches, and combinations thereof. In someembodiments, the diluent is selected from the group consisting oflactose, mannitol, isomalt, microcrystalline cellulose, dicalciumphosphate, and combinations thereof. In some embodiments, the diluent islactose.

In some embodiments, the weight percent of the diluent in the dosageform is from about 70 weight percent to about 99 weight percent. In someembodiments, the weight percent of the diluent in the dosage form isfrom about 80 weight percent to about 99 weight percent. In someembodiments, the weight percent of the diluent in the dosage form isfrom about 85 weight percent to about 99 weight percent. In certain ofthe foregoing embodiments, the diluent is selected from the groupconsisting of lactose, mannitol, isomalt, and combinations thereof. As anon-limiting example, the diluent can be lactose.

(iii) Binder

In some embodiments, the dosage form further comprises apharmaceutically acceptable binder (e.g., polymeric binder). Suitablebinders for use in the disclosed dosage forms include, but are notlimited to, celluloses, such as hydroxypropyl methylcellulose (e.g.,Hypromellose E5 (Premium LV)), hydroxypropyl ethylcellulose, andhydroxypropyl cellulose, and other pharmaceutically acceptablesubstances with cohesive properties. In some embodiments, the binder isselected from the group consisting of hydroxymethylpropylcellulose,hydroxyethylpropylcellulose, and hydroxypropylcellulose. In someembodiments, the binder is hydroxypropyl methylcellulose. In someembodiments, the binder is hydroxypropylcellulose. In some embodiments,the binder is hydroxyethylpropylcellulose.

In some embodiments, the dosage form further comprises apharmaceutically acceptable binder; and the weight percent of the binderin the dosage form is from about 1.0 weight percent to about 10.0 weightpercent. In some embodiments, the weight percent of the binder in thedosage form is from about 1.0 weight percent to about 8.0 weightpercent. In some embodiments, the weight percent of the binder in thedosage form is from about 1.0 weight percent to about 5.0 weightpercent. In certain of the foregoing embodiments, the binder is apolymeric binder selected from the group consisting ofhydroxymethylpropylcellulose, hydroxyethylpropylcellulose, andhydroxypropylcellulose

In some embodiments, the dosage form further comprises apharmaceutically acceptable binder; and the weight to weight ratio ofthe binder to atrasentan, or pharmaceutically acceptable salt thereof,is from about 2:1 to about 25:1 on an atrasentan free base equivalentweight basis. In some embodiments, the weight to weight ratio of thebinder to the atrasentan, or pharmaceutically acceptable salt thereof,is from about 1:1 to about 20:1 on an atrasentan free base equivalentweight basis. In some embodiments, the weight to weight ratio of thebinder to the atrasentan, or pharmaceutically acceptable salt thereof,is from about 1:1 to about 15:1 on an atrasentan free base equivalentweight basis. In certain embodiments, the binder is a polymeric binderselected from the group consisting of hydroxymethylpropylcellulose,hydroxyethylpropylcellulose, and hydroxypropylcellulose;

(iv) Disintegrant

In some embodiments, the dosage form optionally comprises apharmaceutically acceptable disintegrant. Suitable disintegrants for usein the disclosed dosage forms include, but are not limited to,cross-linked polyvinyl pyrrolidone (such as POLYPLASDONE™ XL), cornstarch, potato starch, maize starch and modified starches (includingsodium starch glycolate), agar-agar, alginic acids, microcrystallinecellulose, sodium croscarmellose, and combinations thereof. In someembodiments, the disintegrant is selected from the group consisting ofcrospovidone, sodium starch glycolate, and sodium croscarmellose. Insome embodiments, the disintegrant is a cross-linked polyvinylpyrrolidone. In some embodiments, the disintegrant is crospovidone.

In some embodiments, the dosage form further comprises apharmaceutically acceptable disintegrant. In certain embodiments, theweight percent of the disintegrant in the dosage form is from about 1.0weight percent to about 10.0 weight percent. In some embodiments, theweight percent of the disintegrant in the dosage form is from about 1.0weight percent to about 6.0 weight percent. In some embodiments, theweight percent of the disintegrant in the dosage form is from about 1.0weight percent to about 4.0 weight percent. In certain of the foregoingembodiments, the disintegrant is crospovidone.

In some embodiments, the dosage form further comprises apharmaceutically acceptable disintegrant and the weight to weight ratioof the disintegrant to the anti-oxidant (e.g., L-cysteine), orpharmaceutically acceptable salt or ester thereof, is from about 60:1 toabout 3:1. In some embodiments, the weight to weight ratio of thedisintegrant to the anti-oxidant (e.g., L-cysteine), or pharmaceuticallyacceptable salt or ester thereof, is from about 50:1 to about 4:1. Insome embodiments, the weight to weight ratio of the disintegrant to theanti-oxidant (e.g., L-cysteine), or pharmaceutically acceptable salt orester thereof, is from about 35:1 to about 5:1.

(v) Additional Excipients

In further embodiments, the dosage form optionally comprises apharmaceutically acceptable lubricant and/or glidant. Suitablelubricants and glidants for use in the disclosed dosage forms include,but are not limited to, silicon dioxide (such as SYLOID® 244FP andAEROSIL® 200), glyceryl behenate (such as COMPRITOL®), talc, stearicacid, solid polyethylene glycols, silica gel and mixtures thereof andother substances with lubricating or gliding properties. In certainembodiments, the lubricant is glyceryl behenate (such as COMPRITOL®). Incertain embodiments, the glidant is silicon dioxide (such as SYLOID®244FP). In certain embodiments, the lubricant is glyceryl behenate andthe glidant is silicon dioxide.

In some embodiments, the dosage form further comprises apharmaceutically acceptable glidant. In another aspect, the weightpercent of the glidant in the dosage form is from about 0.1 weightpercent to about 1.5 weight percent. In some embodiments, the weightpercent of the glidant in the dosage form is from about 0.1 weightpercent to about 1.0 weight percent. In some embodiments, the weightpercent of the glidant in the dosage form is from about 0.1 weightpercent to about 0.8 weight percent. In some embodiments, the glidant issilicon dioxide.

In some embodiments, the dosage form further comprises apharmaceutically acceptable lubricant. In some embodiments, the dosageform further comprises a pharmaceutically acceptable, hydrophobiclubricant. In some embodiments, the weight percent of the lubricant inthe dosage form is from about 0.05 weight percent to about 5.0 weightpercent. In some embodiments, the weight percent of the lubricant in thedosage form is from about 0.2 weight percent to about 3.0 weightpercent. In some embodiments, the weight percent of the lubricant in thedosage form is from about 0.5 weight percent to about 2.0 weightpercent. In certain embodiments, the lubricant is glyceryl behenate.

In some embodiments, the dosage form further comprises a disintegrant, aglidant, and a lubricant.

(vi) Anti-Oxidant

Suitable anti-oxidants for use in the disclosed dosage forms includeanti-oxidants that function as reducing agents and are oxidized topharmaceutically acceptable reduced products in the dosage form. In someembodiments, the anti-oxidant has an oxidation reduction potential lessthan the oxidation reduction potential of atrasentan (i.e., an oxidationreduction potential less than about 900 mV) and greater than about 550mV. In some embodiments, the anti-oxidant has an oxidation reductionpotential less than about 550 mV. In some embodiments, the anti-oxidanthas an oxidation reduction potential from about 1 mV to about 550 mV. Insome embodiments, the solubility of the anti-oxidant in water at about25° C. is greater than about 24 mg/mL. In some embodiments, theanti-oxidant is an amino acid, or a pharmaceutically acceptable salt orester thereof. In some embodiments, the anti-oxidant is cysteine. Insome embodiments, the anti-oxidant is L-cysteine, or a pharmaceuticallyacceptable salt or ester thereof. In some embodiments, the anti-oxidantis selected from the group consisting of L-cysteine hydrochloridemonohydrate, L-cysteine hydrochloride anhydrate, and L-cysteine ethylester. In some embodiments, the dosage form comprises L-cysteinehydrochloride monohydrate.

In some embodiments, the weight percent of the anti-oxidant in thedosage form is from about 0.05 weight percent to about 1.0 weightpercent. In some embodiments, the weight percent of the anti-oxidant inthe dosage form is from about 0.07 weight percent to about 0.7 weightpercent. In some embodiments, the weight percent of the anti-oxidant inthe dosage form is from about 0.09 weight percent to about 0.5 weightpercent.

In some embodiments, the molar ratio of the anti-oxidant to atrasentan,or pharmaceutically acceptable salt thereof, is from about 10:1 to about1:10. In some embodiments, the molar ratio of the anti-oxidant toatrasentan, or pharmaceutically acceptable salt thereof, in the dosageform is from about 5:1 to about 1:5. In some embodiments, the molarratio of the anti-oxidant to atrasentan, or pharmaceutically acceptablesalt thereof, is from about 2:1 to about 1:2. In some embodiments, themolar ratio of the anti-oxidant to atrasentan, or pharmaceuticallyacceptable salt thereof, is about 1:1.

In some embodiments, the anti-oxidant is L-cysteine, or apharmaceutically acceptable salt thereof. In certain embodiments, theweight percent of the L-cysteine, or pharmaceutically acceptable salt orester thereof, in the dosage form is from about 0.05 weight percent toabout 1.0 weight percent. In certain embodiments, the weight percent ofthe L-cysteine, or pharmaceutically acceptable salt or ester thereof, inthe dosage form is from about 0.07 weight percent to about 0.7 weightpercent. In certain embodiments, the weight percent of the L-cysteine,or pharmaceutically acceptable salt or ester thereof, in the dosage formis from about 0.09 weight percent to about 0.5 weight percent.

In certain embodiments, the molar ratio of the L-cysteine, orpharmaceutically acceptable salt or ester thereof, to atrasentan, orpharmaceutically acceptable salt thereof, in the dosage form is fromabout 10:1 to about 1:10. In certain embodiments, the molar ratio of theL-cysteine, or pharmaceutically acceptable salt or ester thereof, toatrasentan, or pharmaceutically acceptable salt thereof, in the dosageform is from about 5:1 to about 1:5. In certain embodiments, the molarratio of the L-cysteine, or pharmaceutically acceptable salt or esterthereof, to atrasentan, or pharmaceutically acceptable salt thereof, isfrom about 2:1 to about 1:2. In certain embodiments, the molar ratio ofthe L-cysteine, or pharmaceutically acceptable salt or ester thereof, toatrasentan, or pharmaceutically acceptable salt thereof, about 1.1.

In certain embodiments, the anti-oxidant is selected from the groupconsisting of L-cysteine hydrochloride monohydrate, L-cysteinehydrochloride anhydrate, and L-cysteine ethyl ester. In someembodiments, the dosage form comprises L-cysteine hydrochloridemonohydrate.

(vii) Additional Embodiments

In some embodiments, the dosage form comprises atrasentan or apharmaceutically acceptable salt thereof and an anti-oxidant. In certainof these embodiments, the anti-oxidant is L-cysteine, orpharmaceutically acceptable salt or ester thereof. In some embodiments,the molar ratio of the anti-oxidant (e.g., L-cysteine, orpharmaceutically acceptable salt or ester thereof) is from about 5:1 toabout 1:5. In certain of the foregoing embodiments, the dosage formfurther comprises a pharmaceutically acceptable polymeric binderselected from the group consisting of hydroxymethylpropylcellulose,hydroxyethylpropylcellulose, and hydroxypropylcellulose; the molar ratioof the anti-oxidant (e.g., L-cysteine, or pharmaceutically acceptablesalt or ester thereof) to atrasentan, or pharmaceutically acceptablesalt thereof, is from about 5:1 to about 1:5; and the weight to weightratio of the binder to atrasentan, or pharmaceutically acceptable saltthereof, is from about 1:1 to about 20:1 on an atrasentan free baseequivalent weight basis. In some embodiments, this dosage form furthercomprises a disintegrant and the weight to weight ratio of thedisintegrant to the anti-oxidant (e.g., L-cysteine, or pharmaceuticallyacceptable salt or ester thereof) is from about 60:1 to about 3:1. Insome embodiments, the weight percent of atrasentan, or pharmaceuticallyacceptable salt thereof, in this dosage form is from about 0.2 weightpercent to about 1.0 weight percent on an atrasentan free baseequivalent weight basis. In some embodiments, this dosage form comprisesfrom about 0.40 mg to about 0.85 mg of atrasentan, or an equivalentamount of a pharmaceutically acceptable salt thereof.

In some embodiments, the dosage form comprises a pharmaceuticallyacceptable polymeric binder selected from the group consisting ofhydroxymethylpropylcellulose, hydroxyethylpropylcellulose, andhydroxypropylcellulose; the molar ratio of the anti-oxidant (e.g.,L-cysteine, or a pharmaceutically acceptable salt or ester thereof), toatrasentan, or pharmaceutically acceptable salt thereof, is from about2:1 to about 1:2; and the weight to weight ratio of the binder toatrasentan, or pharmaceutically acceptable salt thereof, is from about1:1 to about 15:1 on an atrasentan free base equivalent weight basis. Insome embodiments, the dosage form further comprises a disintegrant andthe weight to weight ratio of the disintegrant to the anti-oxidant(e.g., L-cysteine, or a pharmaceutically acceptable salt or esterthereof), is from about 50:1 to about 4:1. In some embodiments, theweight percent of atrasentan, or pharmaceutically acceptable saltthereof, in this dosage form is from about 0.2 weight percent to about1.0 weight percent on an atrasentan free base equivalent weight basis.In some embodiments, this dosage form comprises from about 0.40 mg toabout 0.85 mg of atrasentan, or an equivalent amount of apharmaceutically acceptable salt thereof.

In some embodiments, the dosage form comprises a pharmaceuticallyacceptable polymeric binder selected from the group consisting ofhydroxymethylpropylcellulose, hydroxyethylpropylcellulose, andhydroxypropylcellulose; the molar ratio of the anti-oxidant (e.g.,L-cysteine, or pharmaceutically acceptable salt or ester thereof), toatrasentan, or pharmaceutically acceptable salt thereof, is about 1:1;and the weight to weight ratio of the binder to atrasentan, orpharmaceutically acceptable salt thereof, is from about 1:1 to about15:1 on an atrasentan free base equivalent weight basis. In someembodiments, this dosage form further comprises a disintegrant and theweight to weight ratio of the disintegrant to the anti-oxidant (e.g.,L-cysteine, or pharmaceutically acceptable salt or ester thereof), isfrom about 35:1 to about 5:1. In some embodiments, the weight percent ofatrasentan, or pharmaceutically acceptable salt thereof, in this dosageform is from about 0.3 weight percent to about 0.8 weight percent on anatrasentan free base equivalent weight basis. In some embodiments, thisdosage form comprises from about 0.40 mg to about 0.85 mg of atrasentan,or an equivalent amount of a pharmaceutically acceptable salt thereof.

In some embodiments, the dosage form comprises a pharmaceuticallyacceptable polymeric binder selected from the group consisting ofhydroxymethylpropylcellulose, hydroxyethylpropylcellulose, andhydroxypropylcellulose; the dosage form comprises from about 0.05 weightpercent to about 1.0 weight percent of the anti-oxidant (e.g.,L-cysteine, or pharmaceutically acceptable salt or ester thereof); andthe dosage form comprises from about 1.0 weight percent to about 10.0weight percent of the binder. In some embodiments, this dosage formfurther comprises a disintegrant and the weight percent of thedisintegrant in the dosage form is from about 1.0 weight percent toabout 10.0 weight percent. In some embodiments, the weight percent ofatrasentan, or pharmaceutically acceptable salt thereof, in this dosageform is from about 0.1 weight percent to about 2.0 weight percent on anatrasentan free base equivalent weight basis. In some embodiments, thisdosage form comprises from about 0.40 mg to about 0.85 mg of atrasentan,or an equivalent amount of a pharmaceutically acceptable salt thereof.

In some embodiments, the dosage form comprises a pharmaceuticallyacceptable polymeric binder selected from the group consisting ofhydroxymethylpropylcellulose, hydroxyethylpropylcellulose, andhydroxypropylcellulose; the dosage form comprises from about 0.07 weightpercent to about 0.70 weight percent of the anti-oxidant (e.g.,L-cysteine, or pharmaceutically acceptable salt or ester thereof); andthe dosage form comprises from about 1.0 weight percent to about 8.0weight percent of the binder. In some embodiments, this dosage formfurther comprises a disintegrant and the weight percent of thedisintegrant in the dosage form is from about 1.0 weight percent toabout 6.0 weight percent. In some embodiments, the weight percent ofatrasentan, or pharmaceutically acceptable salt thereof, in this dosageform is from about 0.2 weight percent to about 1.0 weight percent on anatrasentan free base equivalent weight basis. In some embodiments, thisdosage form comprises from about 0.40 mg to about 0.85 mg of atrasentan,or an equivalent amount of a pharmaceutically acceptable salt thereof.

In some embodiments, the dosage form comprises a pharmaceuticallyacceptable polymeric binder selected from the group consisting ofhydroxymethylpropylcellulose, hydroxyethylpropylcellulose, andhydroxypropylcellulose; the dosage form comprises from about 0.09 weightpercent to about 0.80 weight percent of the anti-oxidant (e.g.,L-cysteine, or pharmaceutically acceptable salt or ester thereof) andthe dosage form comprises from about 1.0 weight percent to about 5.0weight percent of the binder. In some embodiments, this dosage formfurther comprises a disintegrant and the weight percent of thedisintegrant in the dosage form is from about 1.0 weight percent toabout 4.0 weight percent. In some embodiments, the weight percent ofatrasentan, or pharmaceutically acceptable salt thereof, in this dosageform is from about 0.3 weight percent to about 0.8 weight percent on anatrasentan free base equivalent weight basis. In some embodiments, thisdosage form comprises from about 0.40 mg to about 0.85 mg of atrasentan,or an equivalent amount of a pharmaceutically acceptable salt thereof.

In some embodiments, the dosage form comprises:

(a) about 0.1 weight percent to about 2.0 weight percent of atrasentan,or pharmaceutically acceptable salt thereof, on an atrasentan free baseequivalent weight basis;(b) about 0.05 weight percent to about 1.0 weight percent of theanti-oxidant (e.g., L-cysteine, or pharmaceutically acceptable salt orester thereof);(c) about 75 weight percent to about 99 weight percent of the diluent;(d) about 1.0 weight percent to about 10.0 weight percent of apharmaceutically acceptable binder;(e) optionally, about 1.0 weight percent to about 10.0 weight percent ofa pharmaceutically acceptable disintegrant;(f) optionally, about 0 weight percent to about 1.5 weight percent of apharmaceutically acceptable glidant; and(g) optionally, about 0 weight percent to about 5.0 weight percent of apharmaceutically acceptable lubricant;wherein the cumulative weight percent for all components of the dosageform equals 100 percent.

In some embodiments, the dosage form comprises:

(a) about 0.1 weight percent to about 2.0 weight percent of atrasentan,or pharmaceutically acceptable salt thereof, on an atrasentan free baseequivalent weight basis;(b) about 0.05 weight percent to about 1.0 weight percent of theanti-oxidant (L-cysteine, or pharmaceutically acceptable salt or esterthereof);(c) about 75 weight percent to about 99 weight percent of the diluent;(d) about 1.0 weight percent to about 10.0 weight percent of apharmaceutically acceptable binder;(e) about 1.0 weight percent to about 10.0 weight percent of apharmaceutically acceptable disintegrant;(f) optionally, about 0 weight percent to about 1.5 weight percent of apharmaceutically acceptable glidant; and(g) optionally, about 0 weight percent to about 5.0 weight percent of apharmaceutically acceptable lubricant;wherein the cumulative weight percent for all components of the dosageform equals 100 percent.

In some embodiments, the dosage form comprises:

(a) about 0.2 weight percent to about 1.0 weight percent of atrasentan,or pharmaceutically acceptable salt thereof, on an atrasentan free baseequivalent weight basis;(b) about 0.07 weight percent to about 0.7 weight percent of theanti-oxidant (L-cysteine, or pharmaceutically acceptable salt or esterthereof);(c) about 82 weight percent to about 99 weight percent of the diluent;(d) about 1.0 weight percent to about 8.0 weight percent of apharmaceutically acceptable binder;(e) optionally, about 1.0 weight percent to about 6.0 weight percent ofa pharmaceutically acceptable disintegrant;(f) optionally, about 0 weight percent to about 1.0 weight percent of apharmaceutically acceptable glidant; and(g) optionally, about 0 weight percent to about 3.0 weight percent of apharmaceutically acceptable lubricant;wherein the cumulative weight percent for all components of the dosageform equals 100 percent.

In some embodiments, the dosage form comprises:

(a) about 0.2 weight percent to about 1.0 weight percent of atrasentan,or pharmaceutically acceptable salt thereof, on an atrasentan free baseequivalent weight basis;(b) about 0.07 weight percent to about 0.70 weight percent ofL-cysteine, or pharmaceutically acceptable salt or ester thereof;(c) about 82 weight percent to about 99 weight percent of the diluent;(d) about 1.0 weight percent to about 8.0 weight percent of apharmaceutically acceptable binder;(e) about 1.0 weight percent to about 6.0 weight percent of apharmaceutically acceptable disintegrant;(f) optionally, about 0 weight percent to about 1.0 weight percent of apharmaceutically acceptable glidant; and(g) optionally, about 0 weight percent to about 3.0 weight percent of apharmaceutically acceptable lubricant;wherein the cumulative weight percent for all components of the dosageform equals 100 percent.

In some embodiments, the dosage form comprises:

(a) about 0.3 weight percent to about 0.8 weight percent of atrasentan,or pharmaceutically acceptable salt thereof, on an atrasentan free baseequivalent weight basis;(b) about 0.09 weight percent to about 0.50 weight percent ofL-cysteine, or pharmaceutically acceptable salt or ester thereof;(c) about 87 weight percent to about 99 weight percent of apharmaceutically acceptable diluent;(d) about 1.0 weight percent to about 5.0 weight percent of apharmaceutically acceptable binder;(e) optionally, about 1.0 weight percent to about 4.0 weight percent ofa pharmaceutically acceptable disintegrant;(f) optionally, about 0 weight percent to about 0.75 weight percent of apharmaceutically acceptable glidant; and(g) optionally, about 0 weight percent to about 2.0 weight percent of apharmaceutically acceptable lubricant;wherein the cumulative weight percent for all components of the dosageform equals 100 percent.

In some embodiments, the dosage form comprises:

(a) about 0.3 weight percent to about 0.8 weight percent of atrasentan,or pharmaceutically acceptable salt thereof, on an atrasentan free baseequivalent weight basis;(b) about 0.09 weight percent to about 0.50 weight percent ofL-cysteine, or pharmaceutically acceptable salt or ester thereof;(c) about 87 weight percent to about 99 weight percent of apharmaceutically acceptable diluent;(d) about 1.0 weight percent to about 5.0 weight percent of apharmaceutically acceptable binder;(e) about 1.0 weight percent to about 4.0 weight percent of apharmaceutically acceptable disintegrant;(f) optionally, about 0 weight percent to about 0.75 weight percent of apharmaceutically acceptable glidant; and(g) optionally, about 0 weight percent to about 2.0 weight percent of apharmaceutically acceptable lubricant;wherein the cumulative weight percent for all components of the dosageform equals 100 percent.

In some embodiments, the dosage form comprises:

(a) about 0.1 weight percent to about 2.0 weight percent of atrasentan,or pharmaceutically acceptable salt thereof, on an atrasentan free baseequivalent weight basis;(b) about 75 weight percent to about 99 weight percent of the diluent;(c) about 1.0 weight percent to about 10.0 weight percent of apharmaceutically acceptable binder;(d) optionally, about 1.0 weight percent to about 10.0 weight percent ofa pharmaceutically acceptable disintegrant;(e) optionally, about 0 weight percent to about 1.5 weight percent of apharmaceutically acceptable glidant; and(f) optionally, about 0 weight percent to about 5.0 weight percent of apharmaceutically acceptable lubricant;wherein the cumulative weight percent for all components of the dosageform equals 100 percent.

In some embodiments, the dosage form comprises:

(a) about 0.1 weight percent to about 2.0 weight percent of atrasentan,or pharmaceutically acceptable salt thereof, on an atrasentan free baseequivalent weight basis;(b) about 75 weight percent to about 99 weight percent of the diluent;(c) about 1.0 weight percent to about 10.0 weight percent of apharmaceutically acceptable binder;(d) about 1.0 weight percent to about 10.0 weight percent of apharmaceutically acceptable disintegrant;(e) optionally, about 0 weight percent to about 1.5 weight percent of apharmaceutically acceptable glidant; and(f) optionally, about 0 weight percent to about 5.0 weight percent of apharmaceutically acceptable lubricant;wherein the cumulative weight percent for all components of the dosageform equals 100 percent.

In some embodiments, the dosage form comprises:

(a) about 0.2 weight percent to about 1.0 weight percent of atrasentan,or pharmaceutically acceptable salt thereof, on an atrasentan free baseequivalent weight basis;(b) about 82 weight percent to about 99 weight percent of the diluent;(c) about 1.0 weight percent to about 8.0 weight percent of apharmaceutically acceptable binder;(d) optionally, about 1.0 weight percent to about 6.0 weight percent ofa pharmaceutically acceptable disintegrant;(e) optionally, about 0 weight percent to about 1.0 weight percent of apharmaceutically acceptable glidant; and(f) optionally, about 0 weight percent to about 3.0 weight percent of apharmaceutically acceptable lubricant;wherein the cumulative weight percent for all components of the dosageform equals 100 percent.

In some embodiments, the dosage form comprises:

(a) about 0.2 weight percent to about 1.0 weight percent of atrasentan,or pharmaceutically acceptable salt thereof, on an atrasentan free baseequivalent weight basis;(b) about 82 weight percent to about 99 weight percent of the diluent;(c) about 1.0 weight percent to about 8.0 weight percent of apharmaceutically acceptable binder;(d) about 1.0 weight percent to about 6.0 weight percent of apharmaceutically acceptable disintegrant;(e) optionally, about 0 weight percent to about 1.0 weight percent of apharmaceutically acceptable glidant; and(f) optionally, about 0 weight percent to about 3.0 weight percent of apharmaceutically acceptable lubricant;wherein the cumulative weight percent for all components of the dosageform equals 100 percent.

In some embodiments, the dosage form comprises:

(a) about 0.3 weight percent to about 0.8 weight percent of atrasentan,or pharmaceutically acceptable salt thereof, on an atrasentan free baseequivalent weight basis;(b) about 87 weight percent to about 99 weight percent of apharmaceutically acceptable diluent;(c) about 1.0 weight percent to about 5.0 weight percent of apharmaceutically acceptable binder;(d) optionally, about 1.0 weight percent to about 4.0 weight percent ofa pharmaceutically acceptable disintegrant;(e) optionally, about 0 weight percent to about 0.75 weight percent of apharmaceutically acceptable glidant; and(f) optionally, about 0 weight percent to about 2.0 weight percent of apharmaceutically acceptable lubricant;wherein the cumulative weight percent for all components of the dosageform equals 100 percent.

In some embodiments, the dosage form comprises:

(a) about 0.3 weight percent to about 0.8 weight percent of atrasentan,or pharmaceutically acceptable salt thereof, on an atrasentan free baseequivalent weight basis;(b) about 87 weight percent to about 99 weight percent of apharmaceutically acceptable diluent;(c) about 1.0 weight percent to about 5.0 weight percent of apharmaceutically acceptable binder;(d) about 1.0 weight percent to about 4.0 weight percent of apharmaceutically acceptable disintegrant;(e) optionally, about 0 weight percent to about 0.75 weight percent of apharmaceutically acceptable glidant; and(f) optionally, about 0 weight percent to about 2.0 weight percent of apharmaceutically acceptable lubricant;wherein the cumulative weight percent for all components of the dosageform equals 100 percent.

In some embodiments, the dosage form satisfies one or more of thefollowing conditions:

(a) the diluent is lactose;(b) the dosage form comprises a pharmaceutically acceptable binder andthe binder is hydroxypropyl methylcellulose;(c) the dosage form comprises a pharmaceutically acceptable disintegrantand the disintegrant is crospovidone;(d) the dosage form comprises a pharmaceutically acceptable glidant andthe glidant is silicon dioxide;(e) the dosage form comprises a pharmaceutically acceptable lubricantand the lubricant is glyceryl behenate.

In some embodiments, the dosage form is a solid pharmaceutical dosageform comprising from about 0.25 mg to about 1.25 mg of the atrasentan ora pharmaceutically acceptable salt thereof (e.g., atrasentanhydrochloride) on an atrasentan parent equivalent weight basis. In someembodiments, the pharmaceutical composition comprises from about 0.40 mgto about 1.00 mg of atrasentan or a pharmaceutically acceptable saltthereof (e.g., atrasentan hydrochloride) on an atrasentan parentequivalent weight basis. In some embodiments, the pharmaceuticalcomposition comprises from about 0.40 mg to about 0.85 mg of atrasentanor a pharmaceutically acceptable salt thereof (e.g., atrasentanhydrochloride) on an atrasentan parent equivalent weight basis. In someembodiments, the pharmaceutical composition comprises from about 0.50 mgof atrasentan or a pharmaceutically acceptable salt thereof (e.g.,atrasentan hydrochloride) on an atrasentan parent equivalent weightbasis. In some embodiments, the pharmaceutical composition comprisesfrom about 0.75 mg of atrasentan or a pharmaceutically acceptable saltthereof (e.g., atrasentan hydrochloride) on an atrasentan parentequivalent weight basis. In certain of the foregoing embodiments, thedosage form is a tablet.

In some embodiments, the dosage form is a tablet. In some embodiments,the tablet has a weight from about 37.5 mg to about 1500 mg. In someembodiments, the tablet has a weight from about 50 mg to about 750 mg.In some embodiments, the tablet has a weight from about 50 mg to about250 mg. In some embodiments, the tablet has a weight from about 75 mg toabout 500 mg. In some embodiments, the tablet has a weight from about 75mg to about 150 mg. In some embodiments, the tablet has a weight fromabout 100 mg to about 250 mg. In some embodiments, the tablet has aweight from about 100 mg to about 230 mg. In some embodiments, thetablet has a water content is below about 10%. In certain embodiments,the tablet has a water content of about 4%-6% (e.g., about 4%-5%).

In general, the tablet optionally can be surrounded or coated with atleast one non-rate-controlling layer. The non-rate-controlling layer canbe formed as a single layer, coating or membrane or a plurality ofsingle layers, coatings or membranes. The functions of thenon-rate-controlling layer can include, for example, providing furtherstability for the atrasentan, serving as a process aid and/or as acosmetic enhancement for the formulation, and/or acting as a maskingagent to reduce any undesired odor associated with the formulation (suchas the odor commonly associated with L-cysteine).

When the dosage form comprises a non-rate-controlling layer, thenon-rate-controlling layer can be made of one or more polymers, as wellas, other ingredients known in the art, such as, but not limited to,plasticizers, pigments/opacifiers, waxes, etc. Examples of polymers thatcan be used include, but are not limited to, hydroxypropylmethylcellulose, hydroxypropyl cellulose, methylcellulose, polyvinylalcohol and polyethylene glycol. Examples of plasticizers that can beused include, but are not limited to, polyethylene glycol(s), glycerin,triacetin, triethyl citrate, diethyl phthalate, L-cysteine, and mineraloils. Examples of pigments/opacifiers that can be used include, but arenot limited to, water soluble dyes (for example, sunset yellow,quinoline yellow, erythrosine, and tartrazine), pigments (for example,aluminum lakes, titanium oxides, iron oxides and talc), and naturalproducts (for example, riboflavin, carotenoids, chlorophyll,anthocyanins, and carmine). An example of a wax that can be usedincludes, but is not limited to, a paraffin wax.

In some embodiments, the dosage form is a tablet coated with apharmaceutically acceptable polymer.

In some embodiments, the dosage form is a capsule.

In some embodiments, the dosage form is packaged in a semi-permeablecontainer. In some embodiments, the semi-permeable container is ablister pack.

In some embodiments, the dosage form is packaged in a substantiallyimpermeable container.

In some embodiments, the dosage form is an immediate release dosageform. In some embodiments, the dosage form is an immediate releasetablet and releases at least about 85% of the atrasentan, orpharmaceutically acceptable salt thereof, within about 45 minutes asdetermined in an in vitro dissolution test conducted using a USPDissolution Apparatus 2 (Paddle Apparatus), a 0.01N hydrochloric aciddissolution medium, and a paddle rotation of 50 RPM. In someembodiments, the dosage form is an immediate release tablet and releasesat least about 75% of the atrasentan, or pharmaceutically acceptablesalt thereof, within about 30 minutes.

In some embodiments, the dosage form comprises less than about 1.0weight percent of total impurities resulting from degradation of theatrasentan, or pharmaceutically acceptable salt thereof, after a storageperiod of six months at about 40° C. and about 75% relative humidity. Insome embodiments, degradation of the atrasentan, or pharmaceuticallyacceptable salt thereof, is analyzed using high-performance liquidchromatography.

In some embodiments, the dosage form comprises less than about 0.6weight percent of any single impurity resulting from degradation of theatrasentan, or pharmaceutically acceptable salt thereof, after a storageperiod of six months at about 40° C. and about 75% relative humidity. Insome embodiments, degradation of the atrasentan, or pharmaceuticallyacceptable salt thereof, is analyzed using high-performance liquidchromatography.

In some embodiments, the dosage form comprises less than about 1.0weight percent of total impurities and less than about 0.6 weightpercent of any single impurity resulting from degradation of theatrasentan, or pharmaceutically acceptable salt thereof, after a storageperiod of six months at about 40° C. and about 75% relative humidity. Insome embodiments, degradation of the atrasentan, or pharmaceuticallyacceptable salt thereof, is analyzed using high-performance liquidchromatography.

In certain embodiments, the dosage form is selected from the groupconsisting of:

Tablet Core Composition Ingredient Weight/Weight % mg/Tablet Atrasentan0.31 0.37^(a) Monohydrochloride Lactose Monohydrate 91.19 109.4(Regular) Hypromellose E5 (Premium 3.00 3.6 IV) Crospovidone 3.50 4.2(Polyplasdone ™ XL) Silicon Dioxide (SYLOID ®) 0.50 0.6 GlycerylBehenate 1.50 1.8 (COMPRITOL ®) Purified Water^(b) n/a n/a Total 100%120 mg Film Coated Table Composition Ingredient Weight/Weight % ^(c)mg/Tablet^(d) PEG1450 3 0.1 Hypromellose E3 (Premium 97 3.5 IV) PurifiedWater n/a n/a ^(a)Atrasentan monohydrochloride salt factor = 1.07 (i.e.,0.35 mg free base × 1.07 = 0.37 mg salt). ^(b)Granulation suspensionmedium. Less than 2% in final product. ^(c)Based on aqueous solution of10% solids. ^(d)Based on a 120 mg tablet weight with a coating weightgain of 3%. Tablet Core Composition Ingredient Weight/Weight % mg/TabletAtrasentan 0.4460 0.5350³ Monohydrochloride Lactose Monohydrate 91.05109.3 (Regular) Hypromellose E5 (Premium 3.000 3.600 IV) Crospovidone3.500 4.200 (Polyplasdone ™ XL) Silicon Dioxide (SYLOID ® 0.500 0.600244FP) Glyceryl Behenate 1.500 1.80 (COMPRITOL ®) Purified Water^(b) n/an/a Total 100% 120 mg Film Coated Table Composition IngredientWeight/Weight % ^(c) mg/Tablet^(d) PEG1450 3 0.1080 Hypromellose E3(Premium 97 3.492 IV) Purified Water n/a n/a Total 100% 123.6 mg^(a)Atrasentan monohydrochloride salt factor = 1.07 (i.e., 0.50 mg freebase × 1.07 = 0.5350 mg salt). ^(b)Granulation suspension medium. Lessthan 2% in final product. ^(c)Based on aqueous solution of 10% solids.^(d)Based on a 120 mg tablet weight with a coating weight gain of 3%.Tablet Core Composition Ingredient Weight/Weight % mg/Tablet Atrasentan0.6690 0.8025^(a) Monohydrochloride Lactose Monohydrate 90.83 109.0(Regular) Hypromellose E5 (Premium 3.000 3.600 IV) Crospovidone 3.5004.200 (Polyplasdone ™ XL) Silicon Dioxide (SYLOID ® 0.500 0.600 244FP)Glyceryl Behenate 1.500 1.80 (COMPRITOL ®) Purified Water^(b) n/a n/aTotal 100% 120 mg Film Coated Table Composition Ingredient Weight/Weight% ^(c) mg/Tablet^(d) PEG1450 3 0.1080 Hypromellose E3 (Premium 97 3.492IV) Purified Water n/a n/a Total 100% 123.6 mg ^(a)Atrasentanmonohydrochloride salt factor = 1.07 (i.e., 0.75 mg free base × 1.07 =0.8025 mg salt). ^(b)Granulation suspension medium. Less than 2% infinal product. ^(c)Based on aqueous solution of 10% solids. ^(d)Based ona 120 mg tablet weight with a coating weight gain of 3%.

In certain embodiments, the dosage form is selected from the groupconsisting of:

Tablet Core Composition Ingredient Weight/Weight % mg/Tablet Atrasentan0.4460 0.5350³ Monohydrochloride Lactose Monohydrate 90.91 109.1(Regular) L-Cysteine Hydrochloride 0.1440 0.1728 MonohydrateHypromellose E5 (Premium 3.000 3.600 IV) Crospovidone 3.500 4.200(Polyplasdone ™ XL) Silicon Dioxide (SYLOID ® 0.500 0.600 244FP)Glyceryl Behenate 1.500 1.800 (COMPRITOL ®) Purified Water^(b) n/a n/aTotal 100% 120 mg Film Coated Table Composition Ingredient Weight/Weight%^(c) mg/Tablet^(d) PEG1450 3 0.1 Hypromellose E3 (Premium 97 3.5 IV)Purified Water n/a n/a Total 100% 123.6 mg ^(a)Atrasentanmonohydrochloride salt factor = 1.07 (i.e., 0.5 mg free base × 1.07 =0.5350 mg salt). ^(b)Granulation suspension medium. Less than 2% infinal product. ^(c)Based on aqueous solution of 10% solids. ^(d)Based ona 120 mg tablet weight with a coating weight gain of 3%. Tablet CoreComposition Ingredient Weight/Weight % mg/Tablet Atrasentan 0.66900.8025³ Monohydrochloride Lactose Monohydrate 90.61 108.7 (Regular)L-Cysteine Hydrochloride 0.216 0.2592 Monohydrate Hypromellose E5(Premium 3.000 3.600 IV) Crospovidone 3.500 4.200 (Polyplasdone ™ XL)Silicon Dioxide (SYLOID ® 0.500 0.600 244FP) Glyceryl Behenate 1.5001.800 (COMPRITOL ®) Purified Water^(b) n/a n/a Total 100% 120 mgIngredient Weight/Weight %^(c) mg/Tablet^(d) PEG1450 3 0.1080Hypromellose E3 (Premium 97 3.492 IV) Purified Water n/a n/a Total 100%123.6 mg ^(a)Atrasentan monohydrochloride salt factor = 1.07 (i.e., 0.75mg free base × 1.07 = 0.8025 mg salt). ^(b)Granulation suspensionmedium. Less than 2% in final product. ^(c)Based on aqueous solution of10% solids. ^(d)Based on a 120 mg tablet weight with a coating weightgain of 3%. Tablet Core Composition Ingredient Weight/Weight % mg/TabletAtrasentan 0.31 0.372^(a) Monohydrochloride Lactose Monohydrate 91.09109.3 (Regular) L-Cysteine Hydrochloride 0.0999 0.120 MonohydrateHypromellose E5 (Premium 3.00 3.6 IV) Crospovidone 3.50 4.2(Polyplasdone ™ XL) Silicon Dioxide (SYLOID ® 0.50 0.60 244FP) GlycerylBehenate 1.50 1.8 (COMPRITOL ®) Purified Water^(b) n/a n/a Total 100%120 mg Film Coated Table Composition Ingredient Weight/Weight %^(c)mg/Tablet^(d) PEG1450 3 0.1 Hypromellose E3 (Premium 97 3.5 IV) PurifiedWater n/a n/a Total 100% 123.6 mg ^(a)Atrasentan monohydrochloride saltfactor = 1.07 (i.e., 0.35 mg free base × 1.07 = 0.37 mg salt).^(b)Granulation suspension medium. Less than 2% in final product.^(c)Based on aqueous solution of 10% solids. ^(d)Based on a 120 mgtablet weight with a coating weight gain of 3%.

The formulations of atrasentan or a pharmaceutically acceptable saltthereof and methods of making the same are further described in U.S.Pat. Nos. 9,364,458 and 10,016,393, each of which is incorporated hereinby reference in its entirety.

F. Dosage and Administration

In some embodiments, a dose of atrasentan or a pharmaceuticallyacceptable salt thereof is between about 0.001 mg and 0.1 mg per kg ofthe subject's body weight (e.g., about 0.001, about 0.002, about 0.003,about 0.004, about 0.005, about 0.006, about 0.007, about 0.008, about0.009, about 0.01, about 0.015, about 0.02, about 0.025, about 0.03,about 0.035, about 0.04, about 0.045, about 0.05, about 0.055, about0.06, about 0.065, about 0.07, about 0.075, about 0.08, about 0.085,about 0.09, about 0.095, or about 0.10 mg per kg, or any value inbetween, of the subject's body weight) of atrasentan or an equivalentamount of a pharmaceutically acceptable salt thereof.

In some embodiments, a dose of atrasentan or a pharmaceuticallyacceptable salt thereof is between about 0.1 mg and 10 mg (e.g. about0.1, about 0.2, about 0.3, about 0.4, about 0.5, about 0.6, about 0.7,about 0.75, about 0.8, about 0.9, about 1.0, about 1.1, about 1.2, about1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9,about 2.0, about 2.5, about 3.0, about 3.5, about 4.0, about 4.5, about5.0, about 5.5, about 6.0, about 6.5, about 7.0, about 7.5, about 8.0,about 8.5, about 9.0, about 9.5, or about 10.0 mg, or any value inbetween) of atrasentan or an equivalent amount of a pharmaceuticallyacceptable salt thereof. In certain embodiments, a dose of atrasentan ora pharmaceutically acceptable salt thereof is about 0.75 mg (e.g., whenadministered once per day) of atrasentan or an equivalent amount of apharmaceutically acceptable salt thereof. In certain embodiments, a doseof atrasentan or a pharmaceutically acceptable salt thereof is about0.25 mg (e.g., when administered once per day) of atrasentan or anequivalent amount of a pharmaceutically acceptable salt thereof. Incertain embodiments, a dose of atrasentan or a pharmaceuticallyacceptable salt thereof is about 0.35 mg (e.g., when administered onceper day) of atrasentan or an equivalent amount of a pharmaceuticallyacceptable salt thereof. In certain embodiments, a dose of atrasentan ora pharmaceutically acceptable salt thereof is about 1.0 mg (e.g., whenadministered once per day) of atrasentan or an equivalent amount of apharmaceutically acceptable salt thereof. In certain embodiments, a doseof atrasentan or a pharmaceutically acceptable salt thereof is about1.25 mg (e.g., when administered once per day) of atrasentan or anequivalent amount of a pharmaceutically acceptable salt thereof. Incertain embodiments, a dose of atrasentan or a pharmaceuticallyacceptable salt thereof is about 1.5 mg (e.g., when administered onceper day) of atrasentan or an equivalent amount of a pharmaceuticallyacceptable salt thereof. In certain embodiments, a dose of atrasentan ora pharmaceutically acceptable salt thereof is about 1.75 mg (e.g., whenadministered once per day) of atrasentan or an equivalent amount of apharmaceutically acceptable salt thereof. In certain of theseembodiments, a dose of atrasentan or a pharmaceutically acceptable saltthereof is 0.75 mg (e.g., 1×0.75 mg tablets; or 1.5×0.50 mg tablets) ofatrasentan or an equivalent amount of a pharmaceutically acceptable saltthereof, administered once per day.

In some embodiments, a dose of atrasentan, or salt or solvate thereof,contains a therapeutically effective amount of atrasentan, or salt orsolvate thereof. In other embodiments, a dose of atrasentan, or salt orsolvate thereof, contains less than a therapeutically effective amountof atrasentan, or salt or solvate thereof, (e.g., when multiple dosesare given in order to achieve the desired clinical or therapeuticeffect).

In some embodiments, the therapeutically effective amount of atrasentan,or a pharmaceutically acceptable salt thereof, is from about 0.20 mg toabout 1.5 mg of atrasentan, or an equivalent amount of apharmaceutically acceptable salt thereof. For example, thetherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof can be about 0.20 mg, about 0.30 mg, about 0.40mg, about 0.50 mg, about 0.60 mg, about 0.70 mg, about 0.80 mg, about0.90 mg, about 1.0 mg, about 1.1 mg, about 1.2 mg, about 1.3 mg, about1.4 mg, or about 1.5 mg of atrasentan or an equivalent amount of apharmaceutically acceptable salt thereof. In certain embodiments, thetherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, is from about 0.25 mg to about 1.25 mg ofatrasentan, or an equivalent amount of a pharmaceutically acceptablesalt thereof. In certain embodiments, the therapeutically effectiveamount of atrasentan, or a pharmaceutically acceptable salt thereof, isfrom about 0.40 mg to about 0.85 mg of atrasentan, or an equivalentamount of a pharmaceutically acceptable salt thereof. For example, thetherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof can be about 0.50 mg, about 0.55 mg, about 0.60mg, about 0.65 mg, about 0.70 mg, about 0.75 mg, about 0.80 mg, or about0.85 mg of atrasentan or an equivalent amount of a pharmaceuticallyacceptable salt thereof. As a non-limiting example, the therapeuticallyeffective amount of atrasentan or pharmaceutically acceptable saltthereof can be about 0.75 mg of atrasentan or an equivalent amount of apharmaceutically acceptable salt thereof.

Atrasentan, or salt or solvate thereof, can be administered by anysuitable route and mode. Suitable routes of administering antibodiesand/or antibody-drug conjugate of the present disclosure are well knownin the art and may be selected by those of ordinary skill in the art. Insome embodiments, atrasentan or a pharmaceutically acceptable saltthereof administered parenterally. Parenteral administration refers tomodes of administration other than enteral and topical administration,usually by injection, and include epidermal, intravenous, intramuscular,intraarterial, intrathecal, intracapsular, intraorbital, intracardiac,intradermal, intraperitoneal, intratendinous, transtracheal,subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid,intraspinal, intracranial, intrathoracic, epidural and intrasternalinjection and infusion. In some embodiments, the route of administrationof atrasentan is intravenous injection or infusion. In some embodiments,the route of administration of atrasentan is intravenous infusion. Insome embodiments, the route of administration of atrasentan isintravenous injection or infusion. In some embodiments, the atrasentanis intravenous infusion. In some embodiments, the route ofadministration of atrasentan is oral.

In one embodiment of the methods or uses or product for uses providedherein, atrasentan is administered to the subject daily, twice daily,three times daily or four times daily. In some embodiments, atrasentanis administered to the subject every other day, once about every week oronce about every three weeks. In some embodiments, atrasentan isadministered to the subject once per day. In some embodiments,atrasentan is administered to the subject twice per day. In someembodiments, atrasentan is administered to the subject at a dose ofabout 0.75 mg once per day. In some embodiments, atrasentan isadministered to the subject at a dose of 0.75 mg once per day. In someembodiments, atrasentan is administered to the subject at a dose ofabout 0.25 mg once per day. In some embodiments, atrasentan isadministered to the subject at a dose of 0.25 mg once per day. In someembodiments, atrasentan is administered to the subject at a dose ofabout 0.35 mg once per day. In some embodiments, atrasentan isadministered to the subject at a dose of 0.35 mg once per day. In someembodiments, atrasentan is administered to the subject at a dose ofabout 0.5 mg once per day. In some embodiments, atrasentan isadministered to the subject at a dose of 0.5 mg once per day. In someembodiments, atrasentan is administered to the subject at a dose ofabout 1.0 mg once per day. In some embodiments, atrasentan isadministered to the subject at a dose of 1.0 mg once per day. In someembodiments, atrasentan is administered to the subject at a dose ofabout 1.75 mg once per day. In some embodiments, atrasentan isadministered to the subject at a dose of 1.75 mg once per day.

G. Combination Therapy

The methods of the present disclosure also contemplate treatmentscomprising administering atrasentan or a pharmaceutically acceptablesalt thereof, as described in any of the embodiments of the disclosure,in combination with one or more additional therapeutic agents (such asan inhibitor of one or more elements of therenin-angiotensin-aldosterone system). Accordingly, atrasentan or apharmaceutically acceptable salt thereof as described anywhere hereincan be administered alone or in combination with one or more additionaltherapeutic agents. When administered in combination with one or moreadditional therapeutic agents, separate dosage forms can be administeredto the subject or a single dosage form comprising both atrasentan, or apharmaceutically acceptable salt thereof, and the additional therapeuticagent(s) can be administered to the subject. If administered as aseparate dosage form, the additional therapeutic agent may beadministered simultaneously with the atrasentan dosage form of thepresent disclosure or sequentially (in either order) with the atrasentandosage form of the present disclosure. Administration of two or moreagents in combination can also be referred to herein as“co-administration.”

Representative additional therapeutic agents include, for example,diuretics, antihypertensive agents, therapeutic agents for diabetes ordiabetic complications, and therapeutic agents for hyperlipidemia.

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more diuretics such ashydrochlorothiazide (such as MICROZIDE™ or ORETIC™), hydroflumethiazide(such as SALURON™), bemetanide (such as BUMEX™), torsemide (such asDEMADEX™), metolazone (such as ZAROXOLYN™), chlorothiazide (such asDIURIL™, ESIDRIX™ or HYDRODIURIL™) triamterene (such as DYRENIUM™),ethacrynic acid (such as EDECRIN™), chlorthalidone (such as HYGROTON™),furosemide (such as LASIX™), indapamide (such as LOZOL™) or amiloride(such as MIDAMOR™ or MODURETIC™).

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more thiazide diuretics, suchas chlorothiazide, chlorthalidone, hydrochlorothiazide,trichlormethiazide, indapamide, or metolazone.

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more loop diuretics, such asbumetanide, ethacrynic acid, furosemide, or torsemide.

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more potassium-sparingdiuretics, such as amiloride, eplerenone, spironolactone, andtriamterene.

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more angiotensin convertingenzyme (ACE) inhibitors such as quinapril (such as ACCUPRIL™),fosinopril, perindopril (such as ACEON™), captopril (such as CAPOTEN™),enalapril (such as VASOTEC™), ENALAPRILAT™, ramipril (such as ALTACE™),cilazapril, delapril, fosenopril (such as MONOPRIL™), zofenopril,indolapril, benazepril (such as LOTENSIN™), lisinopril (such asPRINIVIL™ or ZESTRIL™), spirapril, trandolapril (such as MAVIK™),perindep, pentopril, moexipril (such as UNIVASC™), pivopril, temocapril,omapatrilat, imidapril, rescinnamine, benazeprilat, fosinoprilat,ramiprilat, perindoprilat, quinaprilat, trandolaprilat, moexiprilat,Quinoline Yellow WS, or cilazaprilat. In certain embodiments, the ACEinhibitor is selected from: quinapril, fosinopril perindopril,captopril, enalapril, enalaprilat, ramipril, cilazapril, delapril,fosenopril, zofenopril, indolapril, benazepril, lisinopril, spirapril,trandolapril, perindep, pentopril, moexipril, rescinnamine, andpivopril.

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more angiotensin II receptorblockers (ARB) such as candesartan (such as ATACAND™), candesartancilexetil, eprosartan (such as TEVETEN™), irbesartan (such as AVEPRO™)losartan (such as COZAAR™), olmesartan, olmesartan medoxomil (such asBENICAR™) tasosartan, telmisartan (such as MICARDIS™) valsartan (such asDIOVAN™), zolasartan, azilsartan medoxomil, F1-6828K, RNH-6270, UR-7198,Way-126227, KRH-594, TAK-536, BRA-657, or TA-606. In certainembodiments, the ARB is selected from: candesartan, candesartancilexetil, eprosartan, irbesartan, losartan, olmesartan, olmesartanmedoxomil, telmisartan, valsartan, azilsartan medoxomil, and BRA-657.

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more calcium channel blockerssuch as nifedipine (such as ADALAT™, ADALAT CC™, or PROCARDIA™),verapamil (such as GALAN™ COVERA-HS™, ISOPTIN SR™, or VERELAN™),diltiazem (such as CARDIZEM™, CARDIZEM CD™, CARDIZEM LA™, CARDIZEM SR™,DILACOR™, TIAMATE™, or TIAZAC™), isradipine (such as DYNACIRC™ orDYNACIRC CR™), amlodipine (such as NORVASC™), felodipine (such asPLENDIL™), nisoldipine (such as SULAR™), bepridil (such as VASCOR™),vatanidipine, clevidipine, lercanidipine, or dilitiazem.

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more renin inhibitors such asaliskiren (such as TEKTURNA™).

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more aldosterone receptorantagonists such as eplerenone (such as INSPRA™) or spironolactone (suchas ALDACTONE™).

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more alpha blockers such asdozazosin (such as CARDURA™) phenoxybenzamine (such as DIBENZYLINE™),terazosin (such as HYTRIN™), CDR1-93/478, or CR-2991.

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more beta blockers such astimolol (such as BLOCARDEN™) carteolol (such as CARTROL™), carvedilol(such as COREG™), nadolol (such as CORGARD™), propranolol (such asINNOPRAN XL™), betaxolol (such as KERLONE™) penbutolol (such asLEVATOL™), metoprolol (such as LOPRESSOR™ or TOPROL-XL™), atenolol (suchas TENORMIN™), pindolol (such as VISKEN™), or bisoprolol.

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more alpha-beta blockers suchas labetalol (such as NORMODYNE™ or TRANDATE™).

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more central antiadrenergicssuch as methyldopa (such as ALDOMET™), clonidine (such as CATAPRES™ orCATAPRES-TTS™), guanfacine (such as TENEX™), or guanabenz (such asWYTENSIN™).

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more glycosides/inotropicagents such as digoxin (such as LANOXIN™).

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more alpha glucosidaseinhibitors, such as miglitol (such as GLYSET™) or acarbose (such asPRECOSE™).

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more biguanides, such asroseiglitazone (such as AVANDAMET™) or metformin (such as GLUCOPHAGE™ orGLUCOPHAGE XR™).

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more insulins, such asHUMALOG™, HUMALOG 50/50™, HUMALOG 75/25™, HUMULIN 50/50™, HUMALIN 75/25™HUMALIN L™, HUMALIN N™, HUMALIN R™, HUMALIN R U-500™, HUMALIN U™ ILETINII LENTE™, ILETIN II NPH™, ILETIN II REGULAR™, LANTUS™, NOVOLIN 70/30™,NOVILIN N™, NOVILIN R™, NOVOLOG™, or VELOSULIN BR™, and EXUBERA™.

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more meglitnides, such asrepaglinide (such as PRANDIN™) or nateglinide (such as STARLIX™).

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more sulfonylureas, such asglimepiride (such as AMARYL™), glyburide (such as DIABETA™, GLYNASEPRESTAB™ or MICRONASE™), or glipizide (such as GLUCOTROL™, or GLUCOTROLXL™)

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more thiazolidinediones, suchas pioglitazone (such as ACTOS™) or rosiglitazone (such as AVANDIA™).

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with niacin or one or more nicotinic acidderivatives, such as NIACOR™, NIASPAN™, NICOLAR™, or SLO-NIACIN™.

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more fabric acid derivatives,such as clofibrate (such as ATROMID-S™), gemfibrozil (such as LOPID™),or fenofibrate (such as TRICOR™).

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more bile acid sequestrants,such as colestipol (such as COLESTID™), cholestyramine (such asLOCHOLEST™, PREVALITE™, QUESTRAN™, or QUESTRAN LIGHT™), or colesevelam(such as WELCHOL™).

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more cholesterol absorptioninhibitors, such as ezetimibe (such as ZETIA™).

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors(statins) such as fluvastatin (such as LESCOL™) atorvastatin (such asLIPITOR™), lovastatin (such as ALTOCOR™ or MEVACOR™) pravastatin (suchas PRAVACHOL™), rosuvastatin (such as CRESTOR™), simvastatin (such asZOCOR™), or pitavastatin.

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more additional agents. Insome embodiments, the one or more additional agents is animmunosuppressant. In some embodiments, the one or more additionalagents are selected from aminopterin, azathioprine, cyclosporin A,D-penicillamine, gold salts, hydroxychloroquine, leflunomide,methotrexate, minocycline, rapamycin, sulfasalazine, tacrolimus (FK506),and pharmaceutically acceptable salts thereof. As a non-limitingexample, the one or more additional agents can be hydroxychloroquine.

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof may be co-administered with one or more additional therapeuticagents selected group the group consisting of SGLT-2 inhibitor (such ascanagliflozin), GR-immunosuppressant (such as budesonide), MASP-2antibodies (such as OMS721), dual ET1A/ARB inhibitors (such assparsentan), B cell modulators (e.g., APRIL modulators such asatacicept, APL-2, and VIS649), SYK inhibitor (such as fosamatinib),complement factor 3 convertase inhibitor (such as LNP023), NRF2activator (such as Bardoxolone), and RNAi therapeutic targeting the C5component of the complement pathway (e.g., cemdisiram).

In some embodiments, the one or more additional agents are SGLT-2inhibitors. In some embodiments, the one or more additional agents is aSGLT-2 inhibitor selected from dapagliflozin, canagliflozin,ipragliflozin, empaglifozin, bexagliflozin, licogliflozin, janagliflozin(XZP-5695), tofogliflozin, ertugliflozin, henagliflozin (SHR-3824),enavogliflozin (DWP-16001), TA-1887(3-(4-cyclopropylbenzyl)-4-fluoro-1-(β-D-glucopyranosyl)-1H-indole),indole-N-glycoside 18 (3-(4-ethylbenzyl)-1 glucopyranosyl)-1H-indole),sotagliflozin, luseogliflozin, sergliflozin etabonate, remogliflozin,remogliflozin etabonate, and T-1095(((2R,3S,4S,5R,6S)-6-(2-(3-(benzofuran-5-yl)propanoyl)-3-hydroxy-5-methylphenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)etabonate). In some embodiments, the one or more additional agents is aSGLT-2 inhibitor selected from bexagliflozin, canagliflozin,dapagliflozin, empagliflozin, ertugliflozin, ipragliflozin,luseogliflozin, remogliflozin, serfliflozin, licofliglozin,sotagliflozin, and tofogliflozin. In some embodiments, the one or moreadditional agents is bexagliflozin. In some embodiments, the one or moreadditional agents is canagliflozin. In some embodiments, the one or moreadditional agents is dapagliflozin. In some embodiments, the one or moreadditional agents is empagliflozin. In some embodiments, the one or moreadditional agents is ertugliflozin. In some embodiments, the one or moreadditional agents is ipragliflozin. In some embodiments, the one or moreadditional agents is luseogliflozin. In some embodiments, the one ormore additional agents is remogliflozin. In some embodiments, the one ormore additional agents is serfliflozin. In some embodiments, the one ormore additional agents is licofliglozin. In some embodiments, the one ormore additional agents is sotagliflozin. In some embodiments, the one ormore additional agents is tofogliflozin. In some embodiments, the SGLT-2inhibitor is dapagliflozin propylene glycol hydrate. In someembodiments, the SGLT-2 inhibitor is canagliflozin hemihydrate.

In some embodiments, the amount of the SGLT-2 inhibitor is from about 1mg to about 350 mg. For example, about 1 mg to about 175 mg, about 175mg to about 350 mg, or about 90 mg to about 260 mg. In some embodiments,the amount of the SGLT-2 inhibitor is from about 85 mg to about 325 mg.In some embodiments, the amount of the SGLT-2 inhibitor is from about 1mg to about 50 mg, about 20 mg to about 70 mg, about 50 mg to about 100mg, about 70 mg to about 120 mg, about 90 mg to about 140 mg, about 110mg to about 160 mg, about 130 mg to about 180 mg, about 150 mg to about200 mg, about 170 mg to about 220 mg, about 190 mg to about 240 mg,about 210 mg to about 260 mg, about 230 mg to about 280 mg, about 250 mgto about 300 mg, about 270 mg to about 320 mg, or about 290 mg to about350 mg. For example, about 100 mg or about 300 mg. In some embodiments,the amount of the SGLT-2 inhibitor is from about 1 to about 15 mg. Forexample, about 1 to about 10 mg or about 5 to about 15 mg. In someembodiments, the amount of the SGLT-2 inhibitor is from 1 mg to about 3mg, about 2 mg to about 4 mg, about 3 mg to about 5 mg, about 4 mg toabout 6 mg, about 5 mg to about 7 mg, about 6 mg to about 8 mg, about 7mg to about 9 mg, about 8 mg to about 10 mg, about 9 mg to about 11 mg,about 10 mg to about 12 mg, about 11 mg to about 13 mg, about 12 mg toabout 14 mg, or about 13 mg to about 15 mg.

In some embodiments, the SGLT-2 inhibitor is canagliflozin. In someembodiments, 100 mg or 300 mg of canagliflozin is administered. In someembodiments, 100 mg or 300 mg of canagliflozin hemihydrate isadministered. In some embodiments, the SGLT-2 inhibitor isdapagliflozin. In some embodiments, the SGLT-2 inhibitor isdapagliflozin propylene glycol hydrate. In some embodiments, 5 mg or 10mg of dapagliflozin is administered. In some embodiments, 5 mg or 10 mgof dapagliflozin propylene glycol hydrate is administered. In someembodiments, the SGLT-2 inhibitor is empagliflozin. In some embodiments,10 mg or 25 mg of empagliflozin is administered. In some embodiments,the SGLT-2 inhibitor is ertugliflozin. In some embodiments, 5 mg or 15mg of ertugliflozin is administered. In some embodiments, the SGLT-2inhibitor is ipragliflozin. In some embodiments, 25 mg or 50 mg ofipragliflozin is administered. In some embodiments, the SGLT-2 inhibitoris bexagliflozin. In some embodiments, 20 mg of bexagliflozin isadministered. In some embodiments, the SGLT-2 inhibitor issotagliflozin. In some embodiments, 200 mg or 400 mg of sotagliflozin isadministered. In some embodiments, the SGLT-2 inhibitor islicogliflozin. In some embodiments, 15 mg, 50 mg, 75 mg or 150 mg oflicogliflozin is administered.

In any of the embodiments described herein, various combinations ofatrasentan, or a pharmaceutically acceptable salt thereof, and a SGLT-2inhibitor, producing an effect, are contemplated. In some embodiments,the effect, for example, any of the beneficial or desired results asdescribed herein, is greater than the sum of the effect observed whenthe same amount of atrasentan, or a pharmaceutically acceptable saltthereof, when co-administered, and the same amount of the SGLT-2inhibitor when co-administered, are administered as a monotherapy. Insome embodiments, the co-administration of atrasentan, or apharmaceutically acceptable salt thereof, and a SGLT-2 inhibitor,produce an effect, for example, a therapeutic effect using a smallerdose of either, or both, of the compounds as a monotherapy. For example,producing a therapeutic effect using a smaller dose of atrasentan, or apharmaceutically acceptable salt thereof, and/or the SGLT-2 inhibitorcompared to the amount used in monotherapy. For example, in someembodiments, the dose of atrasentan, or a pharmaceutically acceptablesalt thereof, administered in combination with a SGLT-2 inhibitor may beabout 50% to about 90% of the dose of atrasentan, or a pharmaceuticallyacceptable salt thereof, administered as a monotherapy to produce thesame therapeutic effect, e.g., any of the beneficial or desired resultsincluding described herein. In some embodiments, the dose of the SGLT-2inhibitor, administered in combination with atrasentan, or apharmaceutically acceptable salt thereof, may be about 50% to about 90%of the dose of the SGLT-2 inhibitor, administered as a monotherapy toproduce the same therapeutic effect, e.g., any of the beneficial ordesired results including described herein. For example, treating IgAnephropathy, decreasing renal inflammation and/or fibrosis, decreasinghematuria, decreasing proteinuria, stabilizing eGFR, decreasing thenumber of IgA-nephropathy associated disease flares, delaying the onsetof ESRD, decreasing fatigue, and reducing activation of a mesangialcell.

In some embodiments, the present disclosure relates to the use ofatrasentan or a pharmaceutically acceptable salt thereof in combinationwith a second therapeutic for treating a condition as described in thevarious embodiments of the disclosure.

In some embodiments, the present disclosure relates to the use ofatrasentan or a pharmaceutically acceptable salt thereof, for treating acondition as described in the various embodiments of the disclosure,wherein the use comprises one or more additional therapeutic agent.

In some embodiments, the present disclosure relates to a pharmaceuticalcomposition comprising atrasentan or a pharmaceutically acceptable saltthereof, and further comprising one or more additional therapeuticagent.

In some embodiments, the one or more additional therapeutic agentinhibits one or more elements of the renin-angiotensin-aldosteronesystem. In some embodiments, the one or more additional therapeuticagent is selected from the group consisting of diuretics, angiotensinconverting enzyme (ACE) inhibitors, angiotensin II receptor (ARB)blockers, calcium channel blockers, renin inhibitors, and aldosteroneantagonists. In certain particular embodiments, the one or moreadditional therapeutic agent is selected from the group consisting ofangiotensin converting enzyme (ACE) inhibitors and angiotensin IIreceptor blockers (ARBs). In certain embodiments, the one or moreadditional therapeutic agent is selected from one or more angiotensinconverting enzyme inhibitors. In certain embodiments, the one or moreadditional therapeutic agent is selected from one or more angiotensin IIreceptor blockers. In certain embodiments, the one or more additionaltherapeutic agent comprises one or more ACE inhibitors and one or moreARBs. For example, the one or more inhibitors of the renin-angiotensinsystem can be ACE inhibitor, ARB, or a combination thereof. For example,the ACE inhibitor can be selected from: quinapril, fosinoprilperindopril, captopril, enalapril, enalaprilat, ramipril, cilazapril,delapril, fosenopril, zofenopril, indolapril, benazepril, lisinopril,spirapril, trandolapril, perindep, pentopril, moexipril, rescinnamine,and pivopril. For example, the ARB can be selected from: candesartan,candesartan cilexetil, eprosartan, irbesartan, losartan, olmesartan,olmesartan medoxomil, telmisartan, valsartan, azilsartan medoxomil, andBRA-657.

In some embodiments, atrasentan or a pharmaceutically acceptable saltthereof, may be co-administered with a SGLT-2 inhibitor and one or moreACE inhibitors and/or one or more ARBs. In some embodiments, atrasentanor a pharmaceutically acceptable salt thereof, may be co-administeredwith a SGLT-2 inhibitor and one or more ACE inhibitors. In someembodiments, atrasentan or a pharmaceutically acceptable salt thereof,may be co-administered with a SGLT-2 inhibitor and one or more ARBs. Insome embodiments, atrasentan or a pharmaceutically acceptable saltthereof, may be co-administered with a SGLT-2 inhibitor, an ACEinhibitor, and an ARB.

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims. All publications, patents, patentapplications, and sequence accession numbers cited herein are herebyincorporated by reference in their entirety for all purposes.

The disclosure will be more fully understood by reference to thefollowing examples. They should not, however, be construed as limitingthe scope of the disclosure. It is understood that the examples andembodiments described herein are for illustrative purposes only and thatvarious modifications or changes in light thereof will be suggested topersons skilled in the art and are to be included within the spirit andpurview of this application and scope of the appended claims.

SELECTED EMBODIMENTS

Embodiment 1: A method of inhibiting mesangial cell activation in asubject having IgA nephropathy, comprising administering atherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, to the subject;

-   -   wherein the subject has not been previously diagnosed with one        or more of diabetic nephropathy, HIV/AIDS, or acute kidney        failure.

Embodiment 2: A method of treating IgA nephropathy, comprisingadministering a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof, to a subject in need thereof;

-   -   wherein the subject has not been previously diagnosed with one        or more of diabetic nephropathy, HIV/AIDS, or acute kidney        failure.

Embodiment 3: A method of treating IgA nephropathy, comprisingadministering a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof, to a subject in need thereof;

-   -   wherein the subject does not have one or more of diabetic        nephropathy, HIV/AIDS, or acute kidney failure.

Embodiment 4: A method of treating IgA nephropathy, comprisingadministering a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof, to a subject in need thereof;

-   -   wherein the subject does not suffer from one or more of diabetic        nephropathy, HIV/AIDS, or acute kidney failure.

Embodiment 5: A method of treating IgA nephropathy, comprisingadministering a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof, to a subject in need thereof;

-   -   wherein the subject has not been previously diagnosed with one        or more of diabetic nephropathy, HIV/AIDS, prostate cancer, or        acute kidney failure.

Embodiment 6: A method of treating IgA nephropathy, comprisingadministering a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof, to a subject in need thereof;

-   -   wherein the subject has not been previously diagnosed with one        or more of diabetic nephropathy, HIV-related nephropathy,        prostate cancer, or acute kidney failure.

Embodiment 7: A method of treating IgA nephropathy, comprisingadministering a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof, to a subject in need thereof;

-   -   wherein the subject has not been previously diagnosed with one        or more of diabetic nephropathy, HIV-related nephropathy, or        acute kidney failure.

Embodiment 8: A method of treating IgA nephropathy, comprisingadministering a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof, to a subject in need thereof;

-   -   wherein the subject is not being treated for one or more of        diabetic nephropathy, HIV-related nephropathy, or acute kidney        failure.

Embodiment 9: A method of treating IgA nephropathy, comprisingadministering a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof, to a subject in need thereof;

-   -   wherein the subject has been determined to have controlled serum        glucose levels;    -   wherein the subject has not been diagnosed with one or more of        HIV-related nephropathy or acute kidney failure.

Embodiment 10: A method of decreasing renal inflammation and/or fibrosisin a subject having IgA nephropathy, comprising administering atherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, to a subject in need thereof.

Embodiment 11: A method of decreasing the occurrence of hematuria in asubject having IgA nephropathy, comprising administering atherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, to a subject in need thereof.

Embodiment 12: A method of stabilizing eGFR in a subject having IgAnephropathy, comprising administering a therapeutically effective amountof atrasentan, or a pharmaceutically acceptable salt thereof, to asubject in need thereof.

Embodiment 13: A method of decreasing the number of IgA-nephropathyassociated disease flares in a subject having IgA nephropathy,comprising administering a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof, to a subjectin need thereof.

Embodiment 14: A method of delaying the onset of ESRD in a subjecthaving IgA nephropathy, comprising administering a therapeuticallyeffective amount of atrasentan, or a pharmaceutically acceptable saltthereof, to a subject in need thereof.

Embodiment 15: A method of decreasing proteinuria in a subject havingIgA nephropathy, comprising administering a therapeutically effectiveamount of atrasentan, or a pharmaceutically acceptable salt thereof, toa subject in need thereof.

Embodiment 16: The method of Embodiment 1, wherein the mesangialactivation is induced by IgA immune complexes.

Embodiment 17: The method of Embodiment 1, wherein the mesangialactivation is associated with the presence of IgA immune complexes.

Embodiment 18: The method of Embodiment 1, wherein the inhibiting ofmesangial cell activation comprises reducing expression and/or activityof one or more biomarkers indicative of mesangial cell proliferation.

Embodiment 19: The method of Embodiment 1, wherein the inhibiting ofmesangial cell activation comprises reducing mesangial cellinflammation.

Embodiment 20: The method of Embodiment 19, wherein reducing mesangialcell inflammation comprises reducing expression and/or activity of oneor more of IL6, MCP1 or other biomarkers indicative of mesangial cellinflammation.

Embodiment 21: The method of Embodiment 19 or 20, wherein reducingmesangial cell inflammation comprises reducing IL-6 signaling.

Embodiment 22: The method of Embodiment 1, wherein the inhibiting ofmesangial cell activation comprises reducing the pro-fibrotic responsein the mesangial cells.

Embodiment 23: The method of Embodiment 22, wherein reducing thepro-fibrotic response in the mesangial cells comprises reducingexpression and/or activity of one or more of TGF, PDGF, CTGF, MMP,TIMPS, or other biomarkers indicative of mesangial cell fibrosis.

Embodiment 24: The method of Embodiment 22 or 23, wherein reducing thepro-fibrotic response in the mesangial cells comprises reducingexpression and/or activity of one or more of ET1, TGF, PDGF, CTGF, MMP,TIMPS, IGF1, DPEP1, ASL, AMN, ALPL, SLC6A19, IL-6, NF-kB, PKC, PI3K,Src, Ras, ERK1/2, Rho, Rac, Akt, mTOR, NAPDH oxidase, MAPK, cPLA2,TNF-α, IL-1, CAM, COX-2, iNOS, JAK, STAT3, PI3K, Akt/PKB, IKKs, IkBs,MAPK, Ras, Raf, MEK, ERK, MCP1, Cntfr, Il1b, Csf1, Il2ra, Map3k8,Pfkfb3, Nr4a1, Gem, Fosl2, Klf4, F3, Nfkbia, Ifit2, Nr4a2, Klf2, Jag1,Dnajb4, Il1b, Spsb1, Btg2, Atf3, Csf1, Trib1, Zbtb10, Btg1, Rhob, Nfat5,Edn1, Rel, Nr4a3, Nfkb1, Serpine1, Ccl20, Perl, Cxcl2, Map3k8, Traf1,Pik3r1, Pdgfra, Nfkbia, Pik3cg, Pla2g4a, Tiam1, and Pdgfb.

Embodiment 25: The method of any one of Embodiments 22-24, whereinreducing the pro-fibrotic response in the mesangial cells comprisesreducing expression and/or activity of one or more of ET1, TGF, PDGF,CTGF, MMP, TIMPS, IGF1, DPEP1, ASL, AMN, ALPL, SLC6A19, IL-6, PKC, PI3K,Src, Ras, ERK1/2, Rho, Rac, Akt, mTOR, NAPDH oxidase, MAPK, cPLA2,TNF-α, IL-1, CAM, COX-2, iNOS, JAK, STAT3, PI3K, Akt/PKB, IKKs, IkBs,NF-kB, MAPK, Ras, Raf, MEK, ERK, and MCP1.

Embodiment 26: The method of any one of Embodiments 22-25, whereinreducing the pro-fibrotic response in the mesangial cells comprisesreducing expression and/or activity of one or more of Cntfr, Il1b, Csf1,Il2ra, Map3k8, Il1r1, Pfkfb3, Nr4a1, Gem, Fosl2, Klf4, F3, Nfkbia,Ifit2, Nr4a2, Klf2, Jag1, Dnajb4, Il1b, Spsb1, Btg2, Atf3, Csf1, Trib1,Zbtb10, Btg1, Rhob, Nfat5, Edn1, Rel, Nr4a3, Nfkb1, Serpine1, Ccl20,Perl, Cxcl2, Map3k8, Traf1, Pik3r1, Pdgfra, Nfkbia, Pik3cg, Pla2g4a,Tiam1, and Pdgfb.

Embodiment 27: The method of any one of Embodiments 22-25, whereinreducing the pro-fibrotic response in the mesangial cells comprisesreducing NF-κB signaling and/or PDGF signaling.

Embodiment 28: The method of any one of Embodiment 23-27, whereinreducing the pro-fibrotic response in the mesangial cells comprisesreducing matrix secretion by mesangial cells.

Embodiment 29: The method of Embodiment 28, wherein reducing matrixsecretion by mesangial cells comprises reducing expression and/oractivity of one or more of excess matrix secretion by mesangial cells.

Embodiment 30: The method of any one of Embodiments 1-29, wherein thesubject is not currently receiving one or more immunosuppressants.

Embodiment 31: The method of any one of Embodiments 1-30, wherein thesubject is also being administered one or more additional agents.

Embodiment 32: The method of Embodiment 31, wherein the one or moreadditional agents are selected from alcineurin inhibitors, proteasomeinhibitors, aminoquinolines, complement inhibitors, B-cell inhibitors,cytotoxic agents, mTOR inhibitors, and steroids.

Embodiment 33: The method of Embodiment 31 or 32, wherein the one ormore additional agents are immunosuppressants.

Embodiment 34: The method of any one of Embodiments 31-33, wherein theone or more additional agents are steroids.

Embodiment 35: The method of Embodiment 34, wherein the steroids areselected from the group consisting of prednisone, dexamethasone,hydrocortisone, ciclosporin, and combinations of any of the foregoing.

Embodiment 36: The method of Embodiment 31 or 32, wherein the one ormore additional agents are aminoquinolines.

Embodiment 37: The method of any one of Embodiments 31, 32, or 36,wherein the one or more additional agents is hydroxychloroquine.

Embodiment 38: The method of any one of Embodiments 31-37, wherein thedosage of the one or more additional agents is decreased after betweenabout 15 days to about 30 days of treatment with atrasentan, or apharmaceutically acceptable salt thereof.

Embodiment 39: The method of any one of Embodiments 33-38, wherein thedosage of the one or more additional agents is decreased by about 25% toabout 100%.

Embodiment 40: The method of any one of Embodiments 33-39, wherein thedosage of the one or more additional agents is decreased by about 50% toabout 100%.

Embodiment 41: The method of any one of Embodiments 33-40, wherein thedosage of the one or more additional agents is decreased by about 75% toabout 100%.

Embodiment 42: A method of decreasing fatigue in a subject having IgAnephropathy, comprising administering a therapeutically effective amountof atrasentan, or a pharmaceutically acceptable salt thereof, to asubject in need thereof;

-   -   wherein the subject has been determined not to suffer from one        or more of diabetic nephropathy, HIV-related nephropathy,        prostate cancer, or acute kidney failure.

Embodiment 43: The method of Embodiment 42, wherein the fatigue isreduced by about 10% to about 20%.

Embodiment 44: The method of Embodiment 42 or 43, wherein the decreasein fatigue comprises a decrease in the score on one or more of theFatigue Severity Scale, the Chalder Fatigue Scale, the FACIT FatigueScale, the Brief Fatigue Inventory, the FACT-F Subscale, Global Vigorand Affect, the May and Kline Adjective Checklist, the Pearson-ByarsFatigue Feeling Checklist, the Rhoten Fatigue Scale, the Schedule ofFatigue and Anergia, or the Checklist Individual Strength.

Embodiment 45: The method of any one of Embodiments 1-44, wherein thesubject is concomitantly receiving an ACE inhibitor, an ARB, a statin, adiuretic, a calcium channel blocker, a beta blocker, an aldosteroneantagonist, fish oil, hydroxychloroquine, or a combination of any of theforegoing.

Embodiment 46: The method of any one of Embodiments 1-45, wherein thesubject is concomitantly receiving an ACE inhibitor, an ARB, or acombination thereof.

Embodiment 47: The method of Embodiment 45 or 46, wherein the statin isselected from: atorvastatin, fluvastatin, lovastatin, pravastatin,rosuvastatin, simvastatin, and pitavastatin.

Embodiment 48: The method of any one of Embodiments 45-47, wherein thediuretic is selected from: hydrochlorothiazide, trichlormethiazide,hydroflumethiazide, quinethazone, metolazone, chlorothiazide,chlorthalidone, indapamide, methyclothiazide bemetanide, torsemide,piretanide, ethacrynic acid, bumetanide, furosemide, triamterene,spironolactone, eplerenone, and amiloride.

Embodiment 49: The method of any one of Embodiments 45-48, wherein theACE inhibitor is selected from: quinapril, fosinopril, perindopril,captopril, enalapril, enalaprilat, ramipril, cilazapril, delapril,fosenopril, zofenopril, indolapril, benazepril, lisinopril, spirapril,trandolapril, perindep, pentopril, moexipril, rescinnamine, andpivopril.

Embodiment 50: The method of any one of Embodiments 45-49, wherein theACE inhibitor is selected from: quinapril, fosinopril, captopril,enalapril, and lisinopril.

Embodiment 51: The method of any one of Embodiments 45-50, wherein theACE inhibitor is quinapril.

Embodiment 52: The method of any one of Embodiments 45-50, wherein theACE inhibitor is fosinopril.

Embodiment 53: The method of any one of Embodiments 45-50, wherein theACE inhibitor is captopril.

Embodiment 54: The method of any one of Embodiments 45-50, wherein theACE inhibitor is enalapril.

Embodiment 55: The method of any one of Embodiments 45-50, wherein theACE inhibitor is lisinopril.

Embodiment 56: The method of any one of Embodiments 45-55, wherein theARB is selected from: candesartan, candesartan cilexetil, eprosartan,irbesartan, losartan, olmesartan, olmesartan medoxomil, telmisartan,valsartan, azilsartan medoxomil, and BRA-657.

Embodiment 57: The method of any one of Embodiments 45-56, wherein theARB is selected from: candesartan, losartan, olmesartan, and valsartan.

Embodiment 58: The method of any one of Embodiments 45-57, wherein theARB is candesartan.

Embodiment 59: The method of any one of Embodiments 45-57, wherein theARB is losartan.

Embodiment 60: The method of any one of Embodiments 45-57, wherein theARB is olmesartan.

Embodiment 61: The method of any one of Embodiments 45-57, wherein theARB is valsartan.

Embodiment 62: The method of any one of Embodiments 45-61, wherein thedosage of an ACE inhibitor, an ARB, a statin, a diuretic, a calciumchannel blocker, a beta blocker, an aldosterone antagonist, fish oil,hydroxychloroquine, or a combination of any of the foregoing, isdecreased after between about 15 days to about 30 days of treatment withatrasentan, or a pharmaceutically acceptable salt thereof.

Embodiment 63: The method of any one of Embodiments 45-62, wherein thedosage is decreased by about 25% to about 100%.

Embodiment 64: The method of any one of Embodiments 45-63, wherein thedosage is decreased by about 50% to about 100%.

Embodiment 65: The method of any one of Embodiments 45-64, wherein thedosage is decreased by about 75% to about 100%.

Embodiment 66: The method of any one of Embodiments 46 or 49-65, whereinthe dosage of an ACE inhibitor, an ARB, and/or a diuretic, is decreasedafter treatment with atrasentan, or a pharmaceutically acceptable saltthereof.

Embodiment 67: The method of any one of Embodiments 46 or 49-66, whereinthe dosage of an ACE inhibitor, an ARB, and/or a diuretic, is decreasedafter between about 15 days to about 30 days of treatment withatrasentan, or a pharmaceutically acceptable salt thereof.

Embodiment 68: The method of any one of Embodiments 46 or 49-67, whereinthe dosage of an ACE inhibitor, an ARB, and/or a diuretic, is decreasedby about 25% to about 100% after treatment with atrasentan, or apharmaceutically acceptable salt thereof.

Embodiment 69: The method of any one of Embodiments 46 or 49-68, whereinthe dosage of an ACE inhibitor, an ARB, and/or a diuretic, is decreasedby about 50% to about 100% after treatment with atrasentan, or apharmaceutically acceptable salt thereof.

Embodiment 70: The method of any one of Embodiments 46 or 49-69, whereinthe dosage of an ACE inhibitor, an ARB, and/or a diuretic, is decreasedby about 75% to about 100% after treatment with atrasentan, or apharmaceutically acceptable salt thereof.

Embodiment 71: The method of Embodiment 66, wherein the dosage of an ACEinhibitor is decreased after treatment with atrasentan, or apharmaceutically acceptable salt thereof.

Embodiment 72: The method of Embodiment 66, wherein the dosage of an ARBis decreased after treatment with atrasentan, or a pharmaceuticallyacceptable salt thereof.

Embodiment 73: The method of Embodiment 66, wherein the dosage of adiuretic is decreased after treatment with atrasentan, or apharmaceutically acceptable salt thereof.

Embodiment 74: The method of any one of Embodiments 1-73, wherein theatrasentan is administered as a pharmaceutically acceptable salt.

Embodiment 75: The method of any one of Embodiments 1-74, wherein theatrasentan is administered as atrasentan hydrochloride or atrasentanmandelate.

Embodiment 76: The method of any one of Embodiments 1-75, wherein theatrasentan is administered as atrasentan hydrochloride.

Embodiment 77: The method of any one of Embodiments 1-76, wherein theatrasentan is administered as the free base.

Embodiment 78: The method of any one of Embodiments 1-77, wherein thesubject is at a high risk of progression to ESRD.

Embodiment 79: The method of any one of Embodiments 1-78, wherein thesubject has been diagnosed with IgA nephropathy.

Embodiment 80: The method of Embodiment 79, wherein the diagnosis of IgAnephropathy comprises a kidney biopsy, detecting anti-glycan antibodies,detecting deposition of IgA-immune complexes in the kidney, or acombination of any of the foregoing.

Embodiment 81: The method of Embodiment 79 or 80, wherein the diagnosisof IgA nephropathy comprises a kidney biopsy.

Embodiment 82: The method of any one of Embodiments 1-81, wherein thesubject is excreting an average of about 0.5 grams or more of protein inthe urine per day for at least about 3 months prior to the firstadministration of atrasentan, or a pharmaceutically acceptable saltthereof.

Embodiment 83: The method of any one of Embodiments 1-82, wherein thesubject is excreting an average of about 0.75 grams or more of proteinin the urine per day for at least about 3 months prior to the firstadministration of atrasentan, or a pharmaceutically acceptable saltthereof.

Embodiment 84: The method of any one of Embodiments 1-82, wherein thesubject is excreting an average of about 1 gram or more of protein inthe urine per day for at least about 3 months prior to the firstadministration of atrasentan, or a pharmaceutically acceptable saltthereof.

Embodiment 85: The method of any one of Embodiments 1-84, wherein thesubject has an average eGFR of about 20 to about 90 mL/min/1.73 m² forat least about 3 months prior to the first administration of atrasentan,or a pharmaceutically acceptable salt thereof.

Embodiment 86: The method of any one of Embodiments 1-85, wherein thesubject has an average eGFR of about 30 to about 90 mL/min/1.73 m² forat least about 3 months prior to the first administration of atrasentan,or a pharmaceutically acceptable salt thereof.

Embodiment 87: The method of any one of Embodiments 1-85, wherein thesubject has an average eGFR of about 20 to about 60 mL/min/1.73 m² forat least about 3 months prior to the first administration of atrasentan,or a pharmaceutically acceptable salt thereof.

Embodiment 88: The method of any one of Embodiments 1-87, wherein thesubject has an average HbA1c of about 4% to about 6% for at least about3 months prior to the first administration of atrasentan, or apharmaceutically acceptable salt thereof.

Embodiment 89: The method of any one of Embodiments 1-88, wherein thesubject has an average fasting blood glucose level of about 125 mg/dL orless for at least about 3 months prior to the first administration ofatrasentan, or a pharmaceutically acceptable salt thereof.

Embodiment 90: The method of any one of Embodiments 1-89, wherein thesubject maintains a potassium level within the normal physiologic range.

Embodiment 91: The method of any one of Embodiments 1-90, wherein thesubject maintains a sodium level within the normal physiologic range.

Embodiment 92: The method of any one of Embodiments 1-91, wherein thesubject has ALT/AST levels during the administration of atrasentan, or apharmaceutically acceptable salt thereof, that are about the same as theALT/AST levels prior to the first administration of atrasentan, or apharmaceutically acceptable salt thereof.

Embodiment 93: The method of any one of Embodiments 1-92, wherein thesubject has bilirubin levels during the administration of atrasentan, ora pharmaceutically acceptable salt thereof, that are about the same asthe bilirubin levels prior to the first administration of atrasentan, ora pharmaceutically acceptable salt thereof.

Embodiment 94: The method of any one of Embodiments 1-93, wherein thefluid retention in the subject is manageable with diuretics.

Embodiment 95: The method of any one of Embodiments 1-94, wherein theamount of proteins in the urine of the subject is reduced by about 20%to about 80% after between about 15 day and about 30 days of treatmentwith atrasentan, or a pharmaceutically acceptable salt thereof.

Embodiment 96: The method of Embodiment 95, wherein the amount ofproteins in the urine of the subject is reduced by about 35% to about80%.

Embodiment 97: The method of any one of Embodiments 1-96, wherein theamount of proteins in the urine of the subject is reduced by about 100mg/dL to about 500 mg/dL after between about 15 days and about 30 daysof treatment with atrasentan, or a pharmaceutically acceptable saltthereof.

Embodiment 98: The method of any one of Embodiments 1-97, wherein theamount of proteins in the urine of the subject is reduced by about 500mg/dL to about 900 mg/dL after between about 15 days and about 30 daysof treatment with atrasentan, or a pharmaceutically acceptable saltthereof.

Embodiment 99: The method of any one of Embodiments 1-98, wherein therisk of the subject developing ESRD is reduced by about 20% to about 99%after between about 6 months and about 24 months of treatment withatrasentan, or a pharmaceutically acceptable salt thereof.

Embodiment 100: The method of any one of Embodiments 1-99, wherein therisk of the subject developing ESRD is reduced by about 20% to about 99%after between about 12 months and about 24 months of treatment withatrasentan, or a pharmaceutically acceptable salt thereof.

Embodiment 101: The method of any one of Embodiments 1-100, wherein theaverage rate of decrease in eGFR is from about 0.75 mL/min/year to about6 mL/min/year for at least about 3 months prior to the firstadministration of atrasentan, or a pharmaceutically acceptable saltthereof.

Embodiment 102: The method of any one of Embodiments 1-101, wherein theaverage rate of decrease in eGFR is from about 3 mL/min/year to about 6mL/min/year for at least about 3 months prior to the firstadministration of atrasentan, or a pharmaceutically acceptable saltthereof.

Embodiment 103: The method of any one of Embodiments 1-102, wherein theaverage rate of decrease in eGFR is from about 4 mL/min/year to about 5mL/min/year for at least about 3 months prior to the firstadministration of atrasentan, or a pharmaceutically acceptable saltthereof.

Embodiment 104: The method of any one of Embodiments 1-103, wherein theaverage rate of decrease in eGFR is reduced by from about 15% to about30% after between about 6 months and about 24 months of treatment withatrasentan, or a pharmaceutically acceptable salt thereof.

Embodiment 105: The method of any one of Embodiments 1-104, wherein thetherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, is from about 0.20 mg to about 1.5 mg ofatrasentan, or an equivalent amount of a pharmaceutically acceptablesalt thereof.

Embodiment 106: The method of any one of Embodiments 1-105, wherein thetherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, is from about 0.25 mg to about 1.25 mg ofatrasentan, or an equivalent amount of a pharmaceutically acceptablesalt thereof.

Embodiment 107: The method of any one of Embodiments 1-106, wherein thetherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, is from about 0.40 mg to about 0.85 mg ofatrasentan, or an equivalent amount of a pharmaceutically acceptablesalt thereof.

Embodiment 108: The method of any one of Embodiments 1-107, wherein thetherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, is about 0.75 mg of atrasentan, or anequivalent amount of a pharmaceutically acceptable salt thereof.

Embodiment 109: The method of any one of Embodiments 1-108, furthercomprising determining expression and/or activity of one or more of ET1,TGF, PDGF, CTGF, MMP, TIMPS, IGF1, DPEP1, ASL, AMN, ALPL, SLC6A19, IL-6,NF-kB, PKC, PI3K, Src, Ras, ERK1/2, Rho, Rac, Akt, mTOR, NAPDH oxidase,MAPK, cPLA2, TNF-α, IL-1, CAM, COX-2, iNOS, JAK, STAT3, PI3K, Akt/PKB,IKKs, IkBs, NF-kB, MAPK, Ras, Raf, MEK, ERK, MCP1, Cntfr, Il1b, Csf1,Il2ra, Map3k8, Il1r1, Pfkfb3, Nr4a1, Gem, Fosl2, Klf4, F3, Nfkbia,Ifit2, Nr4a2, Klf2, Jag1, Dnajb4, Il1b, Spsb1, Btg2, Atf3, Csf1, Trib1,Zbtb10, Btg1, Rhob, Nfat5, Edn1, Rel, Nr4a3, Nfkb1, Serpine1, Ccl20,Perl, Cxcl2, Map3k8, Traf1, Pik3r1, Pdgfra, Nfkbia, Pik3cg, Pla2g4a,Tiam1, and Pdgfb.

Embodiment 110: The method of any one of Embodiments 1-109, furthercomprising determining expression and/or activity of one or more of ET1,TGF, PDGF, CTGF, MMP, TIMPS, IGF1, DPEP1, ASL, AMN, ALPL, SLC6A19, IL-6,NF-kB, PKC, PI3K, Src, Ras, ERK1/2, Rho, Rac, Akt, mTOR, NAPDH oxidase,MAPK, cPLA2, TNF-α, IL-1, CAM, COX-2, iNOS, JAK, STAT3, PI3K, Akt/PKB,IKKs, IkBs, NF-kB, MAPK, Ras, Raf, MEK, ERK, and MCP1.

Embodiment 111: The method of any one of Embodiments 1-110, furthercomprising determining expression and/or activity of one or more ofCntfr, Il1b, Csf1, Il2ra, Map3k8, Pfkfb3, Nr4a1, Gem, Fosl2, Klf4, F3,Nfkbia, Ifit2, Nr4a2, Klf2, Jag1, Dnajb4, Il1b, Spsb1, Btg2, Atf3, Csf1,Trib1, Zbtb10, Btg1, Rhob, Nfat5, Edn1, Rel, Nr4a3, Nfkb1, Serpine1,Ccl20, Perl, Cxcl2, Map3k8, Traf1, Pik3r1, Pdgfra, Nfkbia, Pik3cg,Pla2g4a, Tiam1, and Pdgfb.

Embodiment 112: The method of any one of Embodiments 1-108, furthercomprising determining expression and/or activity of one or more of ET1,TGF, PDGF, CTGF, MMP, TIMPS, IGF1, DPEP1, ASL, AMN, ALPL, SLC6A19,NF-kB, and IL6 in the subject.

Embodiment 113: The method of Embodiment 109, wherein determining theexpression and/or activity is performed prior to administration of atherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof.

Embodiment 114: The method of Embodiment 109, wherein determining theexpression and/or activity is performed after administration of atherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof.

Embodiment 115: The method of any one of Embodiments 1-114, furthercomprising administering a therapeutically effective amount of a SGLT-2inhibitor.

Embodiment 116: The method of Embodiment 115, wherein the SGLT-2inhibitor is selected from dapagliflozin, canagliflozin, ipragliflozin,empaglifozin, bexagliflozin, licogliflozin, janagliflozin (XZP-5695),tofogliflozin, ertugliflozin, henagliflozin (SHR-3824), enavogliflozin(DWP-16001), TA-1887(3-(4-cyclopropylbenzyl)-4-fluoro-1-(β-D-glucopyranosyl)-1H-indole),indole-N-glycoside 18 (3-(4-ethylbenzyl)-1 glucopyranosyl)-1H-indole),sotagliflozin, luseogliflozin, sergliflozin etabonate, remogliflozin,remogliflozin etabonate, and T-1095(((2R,3S,4S,5R,6S)-6-(2-(3-(benzofuran-5-yl)propanoyl)-3-hydroxy-5-methylphenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)etabonate).

Embodiment 117: The method of Embodiment 115 or 116, wherein the SGLT-2inhibitor is selected from bexagliflozin, canagliflozin, dapagliflozin,empagliflozin, ertugliflozin, ipragliflozin, luseogliflozin,remogliflozin, serfliflozin, licofliglozin, sotagliflozin, andtofogliflozin.

Embodiment 118: The method of any one of Embodiments 115-117, whereinthe SGLT-2 inhibitor is canagliflozin, dapagliflozin, empagliflozin, orertugliflozin.

Embodiment 119: The method of any one of Embodiments 115-117, whereinthe SGLT-2 inhibitor is canagliflozin.

Embodiment 120: The method of any one of Embodiments 115-117, whereinthe SGLT-2 inhibitor is dapagliflozin.

Embodiment 121: The method of any one of Embodiments 115-117, whereinthe SGLT-2 inhibitor is empagliflozin.

Embodiment 122: The method of any one of Embodiments 115-117, whereinthe SGLT-2 inhibitor is ertugliflozin.

Embodiment 123: The method of any one of Embodiments 115-122, whereinthe subject is administered a SGLT-2 inhibitor and one or more ACEinhibitors and/or one or more ARBs.

Embodiment 124: The method of any one of Embodiments 115-123, whereinthe subject is administered a SGLT-2 inhibitor and one or more ACEinhibitors.

Embodiment 125: The method of any one of Embodiments 115-123, whereinthe subject is administered a SGLT-2 inhibitor and one or more ARBs.

Embodiment 126: The method of any one of Embodiments 115-123, whereinthe subject is administered a SGLT-2 inhibitor and an ACE inhibitor.

Embodiment 127: The method of any one of Embodiments 115-123, whereinthe subject is administered a SGLT-2 inhibitor and an ARB.

Embodiment 128: The method of any one of Embodiments 115-123, whereinthe subject is administered a SGLT-2 inhibitor, an ACE inhibitor, and anARB.

Embodiment 129: A method of inhibiting mesangial cell activation,comprising contacting a mesangial cell with an effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof.

Embodiment 130: The method of Embodiment 129, wherein the mesangialactivation is induced by IgA immune complexes.

Embodiment 131: The method of Embodiment 129, wherein the mesangialactivation is associated with the presence of IgA immune complexes.

Embodiment 132: The method of Embodiment 129, wherein the inhibiting ofmesangial cell activation comprises reducing expression and/or activityof one or more biomarkers indicative of mesangial cell proliferation.

Embodiment 133: The method of Embodiment 129, wherein the inhibiting ofmesangial cell activation comprises reducing mesangial cellinflammation.

Embodiment 134: The method of Embodiment 133, wherein reducing mesangialcell inflammation comprises reducing expression and/or activity of oneor more of IL6, MCP1 or other biomarkers indicative of mesangial cellinflammation.

Embodiment 135: The method of Embodiment 134, wherein reducing mesangialcell inflammation comprises reducing IL-6 signaling.

Embodiment 136: The method of Embodiment 129, wherein the inhibiting ofmesangial cell activation comprises reducing the pro-fibrotic responsein the mesangial cells.

Embodiment 137: The method of Embodiment 136, wherein reducing thepro-fibrotic response in the mesangial cells comprises reducingexpression and/or activity of one or more of TGF, PDGF, CTGF, MMP,TIMPS, or other biomarkers indicative of mesangial cell fibrosis.

Embodiment 138: The method of Embodiment 136 or 137, wherein reducingthe pro-fibrotic response in the mesangial cells comprises reducingexpression and/or activity of one or more of ET1, TGF, PDGF, CTGF, MMP,TIMPS, IGF1, DPEP1, ASL, AMN, ALPL, SLC6A19, IL-6, NF-kB, PKC, PI3K,Src, Ras, ERK1/2, Rho, Rac, Akt, mTOR, NAPDH oxidase, MAPK, cPLA2,TNF-α, IL-1, CAM, COX-2, iNOS, JAK, STAT3, PI3K, Akt/PKB, IKKs, IkBs,NF-kB, MAPK, Ras, Raf, MEK, ERK, MCP1, Cntfr, Il1b, Csf1, Il2ra, Map3k8,Il1r1, Pfkfb3, Nr4a1, Gem, Fosl2, Klf4, F3, Nfkbia, Ifit2, Nr4a2, Klf2,Jag1, Dnajb4, Il1b, Spsb1, Btg2, Atf3, Csf1, Trib1, Zbtb10, Btg1, Rhob,Nfat5, Edn1, Rel, Nr4a3, Nfkb1, Serpine1, Ccl20, Perl, Cxcl2, Map3k8,Traf1, Pik3r1, Pdgfra, Nfkbia, Pik3cg, Pla2g4a, Tiam1, and Pdgfb.

Embodiment 139: The method of Embodiment 136 or 137, wherein reducingthe pro-fibrotic response in the mesangial cells comprises reducingexpression and/or activity of one or more of ET1, TGF, PDGF, CTGF, MMP,TIMPS, IGF1, DPEP1, ASL, AMN, ALPL, SLC6A19, IL-6, NF-kB, PKC, PI3K,Src, Ras, ERK1/2, Rho, Rac, Akt, mTOR, NAPDH oxidase, MAPK, cPLA2,TNF-α, IL-1, CAM, COX-2, iNOS, JAK, STAT3, PI3K, Akt/PKB, IKKs, IkBs,NF-kB, MAPK, Ras, Raf, MEK, ERK, and MCP1.

Embodiment 140: The method of Embodiment 136 or 137, wherein reducingthe pro-fibrotic response in the mesangial cells comprises reducingexpression and/or activity of one or more of Cntfr, Il1b, Csf1, Il2ra,Map3k8, Il1r1, Pfkfb3, Nr4a1, Gem, Fosl2, Klf4, F3, Nfkbia, Ifit2,Nr4a2, Klf2, Jag1, Dnajb4, Il1b, Spsb1, Btg2, Atf3, Csf1, Trib1, Zbtb10,Btg1, Rhob, Nfat5, Edn1, Rel, Nr4a3, Nfkb1, Serpine1, Ccl20, Perl,Cxcl2, Map3k8, Traf1, Pik3r1, Pdgfra, Nfkbia, Pik3cg, Pla2g4a, Tiam1,and Pdgfb.

Embodiment 141: The method of Embodiment 136 or 137, wherein reducingthe pro-fibrotic response in the mesangial cells comprises reducingNF-κB signaling and/or PDGF signaling.

Embodiment 142: The method of any one of Embodiment 136-141, whereinreducing the pro-fibrotic response in the mesangial cells comprisesreducing matrix secretion by mesangial cells.

Embodiment 143: The method of Embodiment 142, wherein reducing matrixsecretion by mesangial cells comprises reducing expression and/oractivity of one or more of excess matrix secretion by mesangial cells.

Embodiment 144: A method of reducing activation of a mesangial cell incontact with an IgA immune complex, comprising contacting a mesangialcell with an effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof.

Embodiment 145: The method of Embodiment 144, wherein the reducingactivation of a mesangial cell comprises reducing expression and/oractivity of one or more biomarkers indicative of mesangial cellproliferation.

Embodiment 146: The method of Embodiment 144, wherein the reducingactivation of a mesangial cell comprises reducing mesangial cellinflammation.

Embodiment 147: The method of Embodiment 146, wherein reducing mesangialcell inflammation comprises reducing expression and/or activity of oneor more of IL6, MCP1, or other biomarkers indicative of mesangial cellinflammation.

Embodiment 148: The method of Embodiment 144, wherein the reducingactivation of a mesangial cell comprises reducing the pro-fibroticresponse in the mesangial cells.

Embodiment 149: The method of Embodiment 148, wherein reducing thepro-fibrotic response in the mesangial cells comprises reducingexpression and/or activity of one or more of TGF, PDGF, CTGF, MMP,TIMPS, or other biomarkers indicative of mesangial cell fibrosis.

Embodiment 150: The method of Embodiment 148 or 149, wherein reducingthe pro-fibrotic response in the mesangial cells comprises reducingexpression and/or activity of one or more of ET1, TGF, PDGF, CTGF, MMP,TIMPS, IGF1, DPEP1, ASL, AMN, ALPL, SLC6A19, IL-6, NF-kB, PKC, PI3K,Src, Ras, ERK1/2, Rho, Rac, Akt, mTOR, NAPDH oxidase, MAPK, cPLA2,TNF-α, IL-1, CAM, COX-2, iNOS, JAK, STAT3, PI3K, Akt/PKB, IKKs, IkBs,NF-kB, MAPK, Ras, Raf, MEK, ERK, MCP1, Cntfr, Il1b, Csf1, Il2ra, Map3k8,Il1r1, Pfkfb3, Nr4a1, Gem, Fosl2, Klf4, F3, Nfkbia, Ifit2, Nr4a2, Klf2,Jag1, Dnajb4, Il1b, Spsb1, Btg2, Atf3, Csf1, Trib1, Zbtb10, Btg1, Rhob,Nfat5, Edn1, Rel, Nr4a3, Nfkb1, Serpine1, Ccl20, Perl, Cxcl2, Map3k8,Traf1, Pik3r1, Pdgfra, Nfkbia, Pik3cg, Pla2g4a, Tiam1, and Pdgfb.

Embodiment 151: The method of Embodiment 148 or 149, wherein reducingthe pro-fibrotic response in the mesangial cells comprises reducingexpression and/or activity of one or more of ET1, TGF, PDGF, CTGF, MMP,TIMPS, IGF1, DPEP1, ASL, AMN, ALPL, SLC6A19, IL-6, NF-kB, PKC, PI3K,Src, Ras, ERK1/2, Rho, Rac, Akt, mTOR, NAPDH oxidase, MAPK, cPLA2,TNF-α, IL-1, CAM, COX-2, iNOS, JAK, STAT3, PI3K, Akt/PKB, IKKs, IkBs,NF-kB, MAPK, Ras, Raf, MEK, ERK, and MCP1.

Embodiment 152: The method of Embodiment 148 or 149, wherein reducingthe pro-fibrotic response in the mesangial cells comprises reducingexpression and/or activity of one or more of Cntfr, Il1b, Csf1, Il2ra,Map3k8, Il1r1, Pfkfb3, Nr4a1, Gem, Fosl2, Klf4, F3, Nfkbia, Ifit2,Nr4a2, Klf2, Jag1, Dnajb4, Il1b, Spsb1, Btg2, Atf3, Csf1, Trib1, Zbtb10,Btg1, Rhob, Nfat5, Edn1, Rel, Nr4a3, Nfkb1, Serpine1, Ccl20, Perl,Cxcl2, Map3k8, Traf1, Pik3r1, Pdgfra, Nfkbia, Pik3cg, Pla2g4a, Tiam1,and Pdgfb.

Embodiment 153: The method of any one of Embodiments 148-152, whereinreducing the pro-fibrotic response in the mesangial cells comprisesreducing matrix secretion by mesangial cells.

Embodiment 154: The method of Embodiment 153, wherein reducing matrixsecretion by mesangial cells comprises reducing expression and/oractivity of one or more biomarkers indicative of excess matrix secretionby mesangial cells.

Embodiment 155: The method of Embodiment 144, wherein the reducingactivation of a mesangial cell comprises reducing undesired mesangialcell migration.

Embodiment 156: The method of any one of Embodiments 129-155, whereinthe contacting occurs in vitro.

Embodiment 157: The method of any one of Embodiments 129-155, whereinthe contacting occurs in vivo.

Embodiment 158: A method of treating IgA nephropathy in a subject inneed thereof, comprising:

-   -   a) determining that the subject has IgA-immune complex        deposition in the kidney; and    -   b) administering a therapeutically effective amount of        atrasentan, or a pharmaceutically acceptable salt thereof, to        the subject.

Embodiment 159: The method of Embodiment 158, wherein the subject hasnot been previously diagnosed with one or more of diabetic nephropathy,HIV/AIDS, or acute kidney failure.

Embodiment 160: The method of Embodiment 158, wherein the subject hasnot been previously diagnosed with HIV-related nephropathy.

Embodiment 161: The method of any one of Embodiments 158-160, whereinthe subject is not currently diagnosed with cancer.

Embodiment 162: The method of any one of Embodiments 158-160, whereinthe subject is not currently being treated for cancer.

Embodiment 163: The method of any one of Embodiments 158-160, whereinthe subject has not been previously diagnosed with cancer.

Embodiment 164: The method of Embodiment 163, wherein the cancer is lungcancer or prostate cancer.

Embodiment 165: A method of treating IgA nephropathy in a subject inneed thereof, comprising:

-   -   a) determining that the subject has elevated levels of mesangial        activation; and    -   b) administering a therapeutically effective amount of        atrasentan, or a pharmaceutically acceptable salt thereof, to        the subject.

Embodiment 166: The method of Embodiment 165, wherein determining ofelevated levels of mesangial activation comprises obtaining a samplefrom the subject and assessing the level of mesangial activation in thesame.

Embodiment 167: The method of Embodiment 166, wherein the sample is akidney biopsy sample.

Embodiment 168: The method of Embodiment 166 or 167, wherein the sampleexhibits elevated levels of one or more of: matrix secretion by themesangial cells, IgA-immune complex deposition, mesangial cellproliferation, and endocapillary cell proliferation.

Embodiment 169: The method of any one of Embodiments 166-168, whereinthe sample exhibits elevated levels of IgA-immune complex deposition.

Embodiment 170: The method of any one of Embodiments 165-169, whereinthe subject has been determined to have proteinuria of at least 1 g/dayin at least two of three consecutive readings over the year prior toadministration of a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof.

Embodiment 171: The method of any one of Embodiments 165-170, whereinthe subject has been administered a maximally tolerated stable dose of aRAS inhibitor for at least 12 weeks prior to administration of atherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof.

Embodiment 172: The method of any one of Embodiments 165-171, whereinthe subject is concurrently administered a maximally tolerated stabledose of a RAS inhibitor and a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof.

Embodiment 173: The method of any one of Embodiments 165-171, whereinthe subject has been determined to have hematuria prior toadministration of a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof.

Embodiment 174: The method of Embodiment 173, wherein the hematuria ismicrohematuria.

Embodiment 175: The method of Embodiment 173, wherein the hematuria isgross hematuria.

Embodiment 176: The method of any one of Embodiments 165-175, whereinthe subject has been determined to have an eGFR of at least 30mL/min/1.73 m² prior to administration of a therapeutically effectiveamount of atrasentan, or a pharmaceutically acceptable salt thereof.

Embodiment 177: The method of any one of Embodiments 165-176, whereinthe subject has not been previously diagnosed with one or more ofdiabetic nephropathy, HIV/AIDS, or acute kidney failure.

Embodiment 178: The method of any one of Embodiments 165-177, whereinthe subject has not been previously diagnosed with HIV-relatednephropathy.

Embodiment 179: The method of any one of Embodiments 165-178, whereinthe subject is not currently diagnosed with cancer.

Embodiment 180: The method of any one of Embodiments 165-178, whereinthe subject is not currently being treated for cancer.

Embodiment 181: The method of any one of Embodiments 165-178, whereinthe subject has not been previously diagnosed with cancer.

Embodiment 182: The method of Embodiment 181, wherein the cancer is lungcancer or prostate cancer.

Embodiment 183: The method of any one of Embodiments 165-182, whereinassessing the level of mesangial activation in the sample comprises oneor more of serum analysis, urinalysis, light microscopy andimmunofluorescence microscopy.

Embodiment 184: A method of treating IgA nephropathy in a subject inneed thereof, comprising:

-   -   a) determining that the subject has elevated levels of        IgA-immune complexes in the kidney; and    -   b) administering a therapeutically effective amount of        atrasentan, or a pharmaceutically acceptable salt thereof, to        the subject.

Embodiment 185: The method of Embodiment 184, wherein determining ofelevated levels of IgA-immune complexes in the kidney comprisesobtaining a kidney biopsy sample from the subject and assessing thelevel of IgA-immune complexes in the same.

Embodiment 186: The method of Embodiment 184 or 185, wherein the sampleexhibits elevated levels of one or more of: matrix secretion by themesangial cells, mesangial cell activation, mesangial cellproliferation, and endocapillary cell proliferation.

Embodiment 187: The method of any one of Embodiments 184-186, whereinthe subject has been determined to have proteinuria of at least 1 g/dayin at least two of three consecutive readings over the year prior toadministration of a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof.

Embodiment 188: The method of any one of Embodiments 184-187, whereinthe subject has been administered a maximally tolerated stable dose of aRAS inhibitor for at least 12 weeks prior to administration of atherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof.

Embodiment 189: The method of any one of Embodiments 184-188, whereinthe subject is concurrently administered a maximally tolerated stabledose of a RAS inhibitor and a therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof.

Embodiment 190: The method of any one of Embodiments 184-189, whereinthe subject has been determined to have hematuria prior toadministration of a therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof.

Embodiment 191: The method of Embodiment 190, wherein the hematuria ismicrohematuria.

Embodiment 192: The method of Embodiment 190, wherein the hematuria isgross hematuria.

Embodiment 193: The method of any one of Embodiments 184-192, whereinthe subject has been determined to have an eGFR of at least 30mL/min/1.73 m² prior to administration of a therapeutically effectiveamount of atrasentan, or a pharmaceutically acceptable salt thereof.

Embodiment 194: The method of any one of Embodiments 184-193, whereinthe subject has not been previously diagnosed with one or more ofdiabetic nephropathy, HIV/AIDS, or acute kidney failure.

Embodiment 195: The method of any one of Embodiments 184-194, whereinthe subject has not been previously diagnosed with HIV-relatednephropathy.

Embodiment 196: The method of any one of Embodiments 184-195, whereinthe subject is not currently diagnosed with cancer.

Embodiment 197: The method of any one of Embodiments 184-196, whereinthe subject is not currently being treated for cancer.

Embodiment 198: The method of any one of Embodiments 184-197, whereinthe subject has not been previously diagnosed with cancer.

Embodiment 199: The method of any one of Embodiments 196-198, whereinthe cancer is lung cancer or prostate cancer.

Embodiment 200: The method of any one of Embodiments 185-199, whereinassessing the level of IgA-immune complexes in the sample comprises oneor more of serum analysis, urinalysis, light microscopy andimmunofluorescence microscopy.

Embodiment 201: The method of any one of Embodiments 158-200, furthercomprising administering a SGLT-2 inhibitor.

Embodiment 202: The method of Embodiment 201, wherein the SGLT-2inhibitor is selected from dapagliflozin, canagliflozin, ipragliflozin,empaglifozin, bexagliflozin, licogliflozin, janagliflozin (XZP-5695),tofogliflozin, ertugliflozin, henagliflozin (SHR-3824), enavogliflozin(DWP-16001), TA-1887(3-(4-cyclopropylbenzyl)-4-fluoro-1-(β-D-glucopyranosyl)-1H-indole),indole-N-glycoside 18 (3-(4-ethylbenzyl)-1 glucopyranosyl)-1H-indole),sotagliflozin, luseogliflozin, sergliflozin etabonate, remogliflozin,remogliflozin etabonate, and T-1095(((2R,3S,4S,5R,6S)-6-(2-(3-(benzofuran-5-yl)propanoyl)-3-hydroxy-5-methylphenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)etabonate).

Embodiment 203: The method of Embodiment 200 or 201, wherein the SGLT-2inhibitor is selected from bexagliflozin, canagliflozin, dapagliflozin,empagliflozin, ertugliflozin, ipragliflozin, luseogliflozin,remogliflozin, serfliflozin, licofliglozin, sotagliflozin, andtofogliflozin.

Embodiment 204: The method of any one of Embodiments 201-203, whereinthe SGLT-2 inhibitor is canagliflozin, dapagliflozin, empagliflozin, orertugliflozin.

Examples Example 1. In Vitro Study Using a Cellular Model of Human IgANephropathy

Primary human mesangial cells in a culture are stimulated withpathogenic dgIgA immune complexes isolated from human IgA nephropathypatients or generated in vitro. Proliferation as well aspro-inflammatory and pro-fibrotic responses to these disease causingimmune complexes are observed in mesangial cells within 48 hours. Thecells are treated with atrasentan or a pharmaceutically acceptable saltthereof (e.g., atrasentan hydrochloride) in an appropriate medium.Changes in proliferation, pro-inflammatory responses, and/orpro-fibrotic responses are measured. The result of this study willindicate the extent to which atrasentan attenuates underlying diseaseprocesses in IgA nephropathy in an in vitro model.

Example 2. A Randomized, Double-Blind, Placebo-Controlled Efficacy andSafety Study of Atrasentan in Patients with IgA Nephropathy

This example describes a Randomized, Double-Blind, Placebo-ControlledEfficacy and Safety Study of Atrasentan on IgA nephropathy patients onrenin angiotensin system (RAS) inhibitors with persistent proteinuriaand risk of rapid renal progression.

Study Overview

This protocol encompasses 2 periods. The first period involves theopen-label treatment of approximately 20-30 patients with IgAnephropathy to confirm the safety and tolerability of atrasentan 0.75 mgdaily over 6 weeks. After all patients have completed at least 6 weeksof treatment, an independent Data Monitoring Committee (DMC) reviews allavailable safety data including adverse events, laboratory studies, andvital signs and makes a recommendation as to whether the safety profileis consistent with prior clinical experience, and safety data supportscontinued evaluation in the larger randomized study. If the DMCrecommends proceeding to the randomized study, the initial open-labelpatients continue to receive open-label treatment for 110 weeks(approximately two years).

The second period involves a double-blind, placebo-controlled study ofapproximately 350-400 patients randomized 1:1 to receive atrasentan 0.75mg daily or a matched placebo for 110 weeks. Patients are followedregularly with blood and urine markers of renal function as well assafety assessments.

Both the initial open-label cohort and the randomized double-blindcohort receive assigned treatment up through week 110. All patientssubsequently discontinue study drug at week 110 and return for a safetyfollow up/study completion visit at week 114. Any patients thatdiscontinue treatment prior to week 114 undergo a safety follow-up visitapproximately four weeks after last dose of study drug. After the safetyfollow-up visit, patients that discontinue treatment prematurely areencouraged to remain on study for quarterly efficacy assessments inorder to provide data for the study endpoints.

Objectives:

The primary objectives of this study are:

(1) To confirm the short-term safety and tolerability of atrasentan 0.75mg daily in IgA nephropathy patients; and

(2) To evaluate the effect of atrasentan on proteinuria levels at week24 compared to placebo.

The secondary objectives are:

(3) To evaluate the effect of atrasentan on slowing estimated glomerularfiltration rate (eGFR) loss between baseline and week 114 (4 weeks postcessation of randomized treatment);

(4) To compare two-year on-treatment rates of change in eGFR betweenatrasentan and placebo (eGFR slope week 6 to 110 of randomizedtreatment); and

(5) To compare the percentage of patients achieving a partial clinicalresponse with proteinuria reduction to less than 1 g/day at week 24between atrasentan and placebo.

The exploratory objectives are:

(6) To evaluate the percentage of patients achieving partial andcomplete responses at each time point between week 6 and 114;

(7) To evaluate quality of life (QOL) of patients receiving atrasentancompared to placebo; and

(8) To evaluate the steady-state pharmacokinetics of atrasentan thatsupport exploratory exposure-response analyses.

Number of Subjects

Approximately 20-30 patients are enrolled into the phase 2 safety periodand approximately 350-400 patients are enrolled into the randomizeddouble-blind period.

Patients who discontinue the initial open-label period for reasons otherthan safety may be replaced at the discretion of the sponsor to ensureadequate safety data review after at least 6 weeks of treatment.

Number of Study Sites: Approximately 120 sites worldwide Criteria forInclusion:

The following are the inclusion criteria. Patients must meet ALL of thefollowing inclusion criteria to be enrolled:

(1) male and female patients aged 18 and older;

(2) biopsy-proven diagnosis of IgA nephropathy not due to secondarycauses;

(3) receiving a maximally tolerated stable dose of a RAS inhibitor forat least 12 weeks prior to screening;

(4) proteinuria of at least 1 g/day at screening and on at least two ofthree consecutive readings over the previous year;

(5) eGFR of at least 30 mL/min/1.73 m² at screening;

(6) willing to abide with highly effective forms of contraception asspecified in the protocol throughout the study; and

(7) willing and able to provide written informed consent and comply withall study visits and study procedures.

Exclusion Criteria

The following are the exclusion criteria; patients must meet NONE of thefollowing exclusion criteria to be enrolled:

(1) concurrent diagnosis of another cause of chronic kidney diseaseincluding diabetic kidney disease, hypertensive kidney disease oranother primary glomerulopathy. A history of well controlledhypertension is acceptable;

(2) presence of cellular glomerular crescents in >25% of glomeruli onkidney biopsy (if biopsy available within 6 months of screening) orclinical suspicion of rapidly progressive glomerulonephritis (RPGN);

(3) history of organ transplantation;

(4) use of systemic immunosuppressant medications (or investigationalagents) for at least 2 weeks in the past 6 months;

(5) blood pressure of >160 systolic or >100 diastolic at screening;

(6) history of heart failure or prior hospital admissions for conditionsrelating to fluid overload such as uncontrolled peripheral edema,pleural effusion, or ascites;

(7) history of clinically significant liver disease and transaminase orbilirubin values more than twice the upper limit of normal;

(8) hemoglobin below 9 g/dL at screening or prior history of bloodtransfusion for anemia within 3 months of screening;

(9) history of malignancy unless cancer free for at least 5 years ornonmelanoma skin cancer not requiring ongoing treatment;

(10) pregnancy, breast feeding, or intent to become pregnant during thestudy period for females;

(11) intent to father a child during the study period for males;

(12) have received any investigational or biologic agent within 1 month(or 5 half-lives of the agent, whichever is longer) prior to screening;

(13) concurrent clinically significant, unstable, or uncontrolledcardiovascular, pulmonary, hepatic, renal, gastrointestinal,genitourinary, haematological, coagulation, immunological,endocrine/metabolic, or other medical disorder that, in the opinion ofthe Investigator, might confound the results of the study or poseadditional risk to the patient by their participation in the study; or

(14) history of an alcohol or illicit drug-related disorder.

Note: Screening of a previously ineligible patient may be repeated withmedical monitor approval.

Test Product(s), Dose, and Mode of Administration:

Patients in the open label period receive atrasentan 0.75 mg once dailyby oral administration. Patients in the randomized double-blind periodreceive either: atrasentan 0.75 mg once daily by oral administration; ormatching placebo once daily by oral administration.

Treatment assignment during the randomized study period is determined byan interactive voice/web response system (IXRS). Patients,investigators, and the sponsor do have access to the assigned studytreatment. In the event of a medical emergency the investigator is ableto receive the treatment assignment through the IXRS system if requiredto provide optimal care of the patient. Randomization is stratified byregion (North America vs all other regions) and baseline urinary proteinlevels at screening (>2 g/day vs<2 g/day).

Duration of Treatment:

Screening Duration: up to 4 weeks

Treatment Duration: 110 weeks

Safety Follow up: 4 weeks

Total study duration for an individual patient: Up to 118 weeks(approximately 2.3 years)

Criteria for Evaluation: Efficacy

The primary efficacy endpoint is the change in proteinuria (urineprotein/creatinine ratio based on 24-hour urine collection) frombaseline to week 24 in atrasentan-treated patients as compared toplacebo-treated patients.

The key secondary efficacy endpoint is the rate of change in eGFR asmeasured through a slope calculated from values at baseline through tothe safety follow up/study completion visit (week 114) inatrasentan-treated patients as compared to placebo-treated patients.

Additional secondary efficacy endpoints include: rate of change in eGFRduring two years on treatment as measured through a chronic slopecalculated from values at week 6 through to week 110 inatrasentan-treated patients as compared to placebo-treated patients, andthe percent of patients achieving partial and complete clinicalresponses.

Criteria for Evaluation: Safety

Safety endpoints include:

(1) frequency and severity of adverse events and serious adverse events;

(2) Frequency and severity of adverse events of special interestincluding events of cardiac failure and/or fluid overload; and

(3) Clinically significant changes in safety labs, ECGs, or physicalexamination findings (including vital signs).

Stopping Criteria

An independent data monitoring committee (DMC) is appointed to monitorthe study. The DMC meets periodically to review the safety data andmakes recommendations regarding continuation, modification, suspensionor termination of the study. The DMC also makes a formal recommendationto proceed to the randomized study following completion of at least 6weeks of treatment for all patients in the open-label period.

The only required study drug stopping criteria is evidence of pregnancyor noncompliance with protocol-specified contraception or pregnancymonitoring. Other study drug stopping criteria may be based oninvestigator and patient discretion that may include fluid overloadsyndromes uncontrolled with diuretics and medical management and anysuspected study drug-related adverse event that represents unacceptabletoxicity.

All patients that stop study drug prior to week 110 are encouraged tocontinue quarterly visits through week 114 for efficacy assessmentsonly.

Criteria for Evaluation: Pharmacokinetics and Pharmacodynamics

The pharmacokinetic and pharmacodynamic endpoints are:

(1) atrasentan plasma levels; and

(2) exploratory biomarkers.

Note: PK sampling is sparse to support development of a population PK.

Statistical Methods Treatment Population Definitions:

-   -   All open-label participants comprise the Open Label Population        for safety analyses and descriptive noncomparative efficacy        analyses    -   All randomised participants who receive any amount of study drug        comprise the Safety Population for safety and exposure analyses    -   All randomised participants comprise the Intent-to-Treat        Population for efficacy analyses    -   All randomised participants who receive any amount of study drug        and who have at least one post-baseline PK sample comprise the        PK Population

Safety and Tolerability:

Continuous safety data are summarised with descriptive statistics(arithmetic mean, standard deviation [SD], median, minimum, and maximum)by treatment group. Categorical safety data are summarised withfrequency counts and percentages by treatment group. Adverse events arecoded using the most current Medical Dictionary for RegulatoryActivities (MedDRA) version available. The number of participantsexperiencing treatment-emergent adverse events as well as maximumseverity and relationship to study drug are summarized.

Laboratory evaluations, vital signs assessments, and ECG parameters aresummarised by treatment group and protocol-specified collection timepoint. A summary of change-from-baseline at each protocol-specified timepoint by treatment group are also presented. Concomitant medications arelisted by participant and coded using the most current World HealthOrganization drug dictionary. Medical history is listed by subject.Further details regarding presentation and analysis of safety data isdetailed in the Statistical Analysis Plan (SAP).

Pharmacokinetics:

Individual steady state atrasentan concentration data is listed andsummarised with descriptive statistics (sample size, arithmetic mean,SD, median, minimum, maximum, geometric mean, and geometric coefficientof variation). These data may be fed into a larger population PK modelof atrasentan. Correlations between atrasentan drug levels and safety orefficacy responses may be explored. Details regarding the statisticalanalyses of pharmacokinetic data are provided in the SAP.

Efficacy Evaluations:

Descriptive statistics are used to evaluate differences in demographicand baseline characteristics. The primary efficacy endpoint is thechange from baseline to each postbaseline visit up to week 24 in urinaryprotein to creatinine ratio as determined from 24-hour urine collectionsamples. The primary analysis is carried out using log transformed dataand a repeated measures analysis using all data collected at baseline,week 12, and week 24. Additional sensitivity analyses are performed onthe primary endpoint to evaluate robustness and the impact of missingdata. Subgroup analyses are performed as specified in the SAP. Followingthe collection and analysis of primary endpoint data at week 24,patients in the randomized study continue to receive their assignedstudy treatment up until study completion at week 114.

Example 3. In Vitro Study Using a Cellular Model of Human IgANephropathy

Primary human mesangial cells in a culture were stimulated withpathogenic galactose deficient (Gd)-IgA containing immune complexesisolated from human IgA nephropathy patients. Proliferation as well aspro-inflammatory and pro-fibrotic responses to these disease causingimmune complexes were observed in mesangial cells within 48-72 hours.The cells were treated with atrasentan and changes in proliferation,pro-inflammatory responses, and/or pro-fibrotic responses were measured.

Specifically, pathogenic IgA-containing immune complexes were isolatedfrom serum from either IgA nephropathy patients or age, gender andethnicity-matched healthy individuals. Frozen serum was purchased fromBioIVT and IgA-containing immune complexes were purified using jacalinaffinity chromatography. In brief, 2 mL of serum was applied to ajacalin-agarose affinity column (ThermoFisher) and IgA was eluted with0.1 M melibiose (Sigma Aldrich). Purified IgA-containing immunecomplexes were concentrated using a 100K Amicon filtration unit and theamount of total IgA and galactose-deficient IgA (Gd-IgA) in the immunecomplexes was measured by ELISA (ThermoFisher and IBL America,respectively). Primary human renal mesangial cells (HRMCs) from a23-year-old male donor were purchased from ScienCell ResearchLaboratories. HRMCs were cultured as recommended by the supplier in MCMmedia containing 2% FBS and proprietary growth factors. Forproliferation and cytokine production experiments, 5,000 cells per wellwere plated in 96 well plates, cultured overnight in complete media,then switched to MCM media containing 0.5% FBS in the absence of growthfactors. Following one day of low serum treatment, media was replacedwith MCM containing 0.5% FBS in the presence of purified immunecomplexes with or without atrasentan for up to 72 hours. See, e.g.,Novak et. al., Kidney International, 67, pp. 504-516; 2005; Novak et.al., Nephrol. Dial. Transplant, 26, pp. 3451-3457; 2011; Liang et. al.,Cell Physiol. Biochem., 36, pp. 1793-1808; 2015; Nguyen et. al., Clin.Kidney J., 12 (2), pp, 232-238; 2019. Mesangial cell activation wasassessed by measuring proliferation with BrdU incorporation (Abcam) andsecretion of inflammatory cytokines including IL-6 was measured by ELISA(R&D Systems).

Compared to the IgA containing serum fractions from healthy controls,HRMCs treated with IgA containing immune complexes purified from threeseparate patients with IgA nephropathy showed increased cellproliferation, a hallmark of mesangial cell activation in response topathogenic IgA immune complexes (FIG. 1A). Atrasentan had no effect onbasal cell proliferation in HRMCs treated with IgA containing serumfractions from healthy controls, however, atrasentan significantlyattenuated the hyperproliferation of HRMCs induced by IgA containingimmune complexes purified from three separate patients with IgAnephropathy (FIG. 1A). The purified immune complexes from normal andIgAN donors contained a mean of 1.9 and 4.4 μg/mL Gd-IgA, respectively(FIG. 1B).

Example 4. Atrasentan Inhibits ET-1 Induced Proliferation and IL-6Production in Primary Human Mesangial Cells

Primary human renal mesangial cells (HRMCs) from a 23-year-old maledonor were purchased from ScienCell Research Laboratories. HRMCs werecultured as recommended by the supplier in MCM media containing 2% FBSand proprietary growth factors (ScienCell). For proliferation andcytokine production experiments, 5,000 cells per well were plated in 96well plates, cultured overnight in complete media, then switched to MCMmedia containing 0.5% FBS in the absence of growth factors. Followingone day of low serum treatment, media was replaced with MCM containing0.5% FBS in the presence or absence of 10 ng/mL ET-1 and a range ofatrasentan concentrations for up to 72 hours. Mesangial cell activationwas assessed by measuring proliferation with bromodeoxyuridine (BrdU)incorporation (Abcam) and secretion of inflammatory cytokines includingIL-6 was measured by ELISA (R&D Systems). IC₅₀ values were calculated inGraphPad Prism using a variable slope four-parameter fit.

ET-1 induced HRMC proliferation at both 48 (FIG. 2A) and 72 hours (FIG.2B). ET-1 induced HRMC proliferation was blocked in aconcentration-dependent manner by atrasentan with IC₅₀ values of 4.2 nMand 50.8 nM after 48 (FIG. 2A) and 72 hours (FIG. 2B) of treatment,respectively. ET-1 also stimulated the production of IL-6 in HRMCs at 48(FIG. 3A) and 72 hours (FIG. 3B), which was inhibited in aconcentration-dependent manner by atrasentan with IC₅₀ values of 1.0 nMand 0.65 nM at 48 and 72 hour timepoints, respectively (FIGS. 3A and3B).

Example 5. A Phase 3, Randomized, Double-Blind, Placebo-Controlled Studyof Atrasentan in Patients with IgA Nephropathy at Risk of ProgressiveLoss of Renal Function (The ALIGN Study)

This example describes a Phase 3, Randomized, Double-Blind,Placebo-Controlled Study of Atrasentan in patients with IgA nephropathy)at risk of progressive loss of renal function despite treatment with amaximally tolerated and stable dose of a RAS inhibitor.

Study Overview

Approximately 320 patients with IgA nephropathy will be randomized 1:1to receive atrasentan 0.75 mg once daily (QD) or a matched placebo for132 weeks. Randomization will be stratified according to region andscreening UPCR levels. Subjects will be evaluated for safety as well aschanges in proteinuria and eGFR from baseline. Total duration for studyparticipation per subject is expected to be up to 140 weeks including 4weeks for screening, 132 weeks of treatment, and 4 weeks follow-up.

An independent data monitoring committee (IDMC) will periodicallyconvene to review unblinded overall safety and emerging efficacyresults. As well, an interim re-estimation of sample size supporting thekey secondary endpoint may result in an increase of enrollment up to 450total subjects

Subjects who complete the study may be eligible to enroll in anextension study to receive open-label treatment with atrasentan under aseparate protocol.

Objectives:

The primary objective of this study is:

(1) To evaluate the effect of atrasentan on proteinuria levels at week24 compared to placebo.

The secondary objectives are:

(2) To evaluate the effect of atrasentan versus placebo on change frombaseline to Week 136 (4 weeks post cessation of randomized treatment) inestimated glomerular filtration rate (eGFR);

(3) To compare 2-year on-treatment rates of change in eGFR betweenatrasentan and placebo (eGFR slope Week 12 to Week 120 of randomizedtreatment); and

(4) To compare the total on-study rates of change in eGFR betweenatrasentan and placebo (eGFR slope from baseline to Week 136).

The exploratory objectives are:

(5) To evaluate quality of life (QOL) of patients receiving atrasentancompared to placebo; and

(6) To evaluate the steady-state pharmacokinetics of atrasentan thatsupport exploratory exposure-response analyses.

Number of Subjects

Approximately 320 patients will be enrolled.

Number of Study Sites: Approximately 150 Sites Worldwide Criteria forInclusion:

The following are the inclusion criteria. Patients must meet all of thefollowing inclusion criteria to be enrolled:

-   -   1. Male and female subjects aged 18 and older at the time of        signing the ICF prior to initiation of any study specific        activities/procedures.    -   2. Biopsy-proven IgAN that, in the opinion of the Investigator,        is not due to secondary causes.        -   Biopsy could have occurred at any point in time prior to            study.        -   A diagnostic report must be available for review by the            Sponsor or designee.    -   3. Receiving a maximally tolerated and optimized dose of a RAS        inhibitor that has been stable for at least 12 weeks prior to        screening.        -   Investigator discretion should be used in determining            maximally tolerated and optimized dose.        -   Subjects who are intolerant to RAS inhibitors are eligible,            but will not exceed ˜5% of total population randomized.    -   4. UPCR ≥1 g/g based on a central laboratory assessment of first        morning void urine collected at screening.    -   5. eGFR of at least 30 mL/min/1.73 m² at screening based on the        CKD-EPI equation.    -   6. Willing to abide with highly effective forms of        contraception, as specified in the protocol, throughout the        study and for 1 month afterward. In WOCBP, use of hormonal        contraceptive agents    -   7. Willing and able to provide written informed consent and        comply with all study visits and study procedures.

Exclusion Criteria

The following are the exclusion criteria; patients must meet none of thefollowing exclusion criteria to be enrolled:

-   1. Concurrent diagnosis of another cause of chronic kidney disease    including diabetic kidney disease or another primary glomerulopathy.-   2. Clinical suspicion of rapidly progressive glomerulonephritis    (RPGN) based on KDIGO guidelines or clinical suspicion of    Henoch-Schonlein Purpura.-   3. Diagnosis of nephrotic syndrome with serum albumin <3 g/dL at    screening.-   4. BNP value of >200 pg/mL at screening.-   5. Platelet count <80,000 per μL at screening-   6. History of organ transplantation (subjects with history of    corneal transplant are not excluded).-   7. Use of systemic immunosuppressant medications including    mycophenolate, azathioprine, cyclosporine, tacrolimus, etc.; use of    herbs such as Tripterygium Wilfordii Hook F, Caulis sinomenii and    Sinomenium acutum; for >2 weeks in the past 3 months. Use of    rituximab within the past 6 months.-   8. Confirmed blood pressure >150 mmHg systolic or >95 mmHg diastolic    based on a mean of 3 measurements obtained at screening.-   9. Known history of heart failure or prior hospital admissions for    conditions relating to fluid overload such as pulmonary edema,    uncontrolled peripheral edema, pleural effusion, or ascites.-   10. Known history of clinically significant liver disease or    transaminase or bilirubin values more than twice the upper limit of    normal. Subjects with treated hepatitis C can be considered for    inclusion into the study upon consultation with the Sponsor's    Medical Monitor (or designee).-   11. Hemoglobin below 9 g/dL at screening or prior history of blood    transfusion for anemia within 3 months of screening.-   12. History of malignancy unless cancer free for at least 5 years or    nonmelanoma skin cancer not requiring ongoing treatment. A subject    with curatively treated cervical carcinoma in situ is eligible for    this study.-   13. Pregnancy, breast feeding, or intent to become pregnant during    the study period and at least 1 month afterward for females.-   14. Intent to father a child or donate sperm during the study period    and at least 1 month afterward for males.-   15. Have received any investigational agent within 1 month (or 5    half-lives of the agent, whichever is longer) prior to screening. If    the investigational agent is a cytotoxic or immunosuppressive agent    then this washout period is 6 months.-   16. Concurrent clinically significant, unstable, or uncontrolled    cardiovascular, pulmonary, hepatic, renal, gastrointestinal,    genitourinary, hematological, coagulation, immunological,    endocrine/metabolic, or other medical disorder that, in the opinion    of the Investigator or Sponsor's Medical Monitor (or designee),    might confound the results of the study or pose additional risk to    the subject by their participation in the study.-   17. History of an alcohol or illicit drug-related disorder within    the past 3 years.

Test Product(s), Dose, and Mode of Administration:

Subjects will receive either atrasentan 0.75 mg once daily by oraladministration or matching placebo once daily by oral administration.

Treatment assignment is determined by an interactive voice/web responsesystem (IXRS). Patients, investigators, and the sponsor do have accessto the assigned study treatment. In the event of a medical emergency theinvestigator is able to receive the treatment assignment through theIXRS system if required to provide optimal care of the patient.Randomization is stratified by region (Asia vs. all other regions) andUPCR levels at screening (?2 g/day vs.<2 g/day).

Duration of Treatment:

Screening Duration: up to 4 weeks

Treatment Duration: 132 weeks

Safety Follow up: 4 weeks

Total study duration for an individual patient: Up to 140 weeks(approximately 2.3 years)

Criteria for Evaluation: Efficacy

The primary efficacy endpoint is the change in proteinuria (urineprotein/creatinine ratio based on 24-hour urine collection) frombaseline to week 24 in atrasentan-treated patients as compared toplacebo-treated patients.

The key secondary endpoint, change from baseline in eGFR for eachsubject, will be determined by the difference in eGFR at Week 136 frombaseline. The eGFR change from baseline will be analyzed using an MMRMmodel.

Additional secondary efficacy endpoints include:

-   -   Rate of change in eGFR during 2 years on treatment as measured        through a chronic slope calculated from values at Week 12        through to Week 120    -   Rate of change in eGFR during the study as measured through a        total slope calculated from values at baseline to Week 136    -   Percent of subjects achieving proteinuria reduction to <1 g/day        at Week 24 and 40% decrease in UPCR from baseline    -   Percent of subjects experiencing at least a 30% reduction in        eGFR or reach ESKD during the study    -   Percent of subjects experiencing at least a 40% reduction in        eGFR or reach ESKD during the study. Secondary endpoints will to        be tested in hierarchical fashion at the final analysis at Week        136 after approximately 320 subjects in the ITT analysis set        complete the Week 136 visit or discontinue from the study.        Secondary analyses will be tested in hierarchical fashion (as        listed above) based on a 2-sided significance level of 0.05.

Criteria for Evaluation: Safety

Safety endpoints include:

(1) frequency and severity of adverse events and serious adverse events;

(2) Frequency and severity of adverse events of special interestincluding events of cardiac failure and/or fluid overload; and

(3) Clinically significant changes in safety labs, ECGs, or physicalexamination findings (including vital signs).

Stopping Criteria

An independent data monitoring committee (DMC) is appointed to monitorthe study. The DMC meets periodically to review the safety data andmakes recommendations regarding continuation, modification, suspensionor termination of the study.

Subjects will be required to permanently discontinue study drug for thefollowing reasons:

-   -   Evidence of pregnancy or noncompliance with protocol-specified        contraception or pregnancy monitoring    -   Chronic dialysis or kidney transplantation for ESKD    -   Other study drug stopping criteria may be based on investigator        and subject discretion that may include fluid overload syndromes        uncontrolled with diuretics and medical management, and any        suspected study drug-related adverse event that represents        unacceptable toxicity.

All subjects who discontinue study drug prior to Week 132 will have anEoT visit at the time of study drug discontinuation, followed by a visit2 weeks after study drug discontinuation and a safety follow up visit 4weeks after study drug discontinuation. Thereafter, subjects shouldcontinue quarterly efficacy assessments for visits through Week 136/EoS.

Criteria for Evaluation: Pharmacokinetics and Pharmacodynamics

The pharmacokinetic and pharmacodynamic endpoints are:

(1) atrasentan plasma levels; and

(2) Proteomic, metabolomic and transcriptomic analysis of the blood andurine for potential biomarkers of IgAN disease activity (such asgalactose deficient IgA (gd-IgA) and gd-IgA auto antibodies) or factorscontributing to the subject's response to atrasentan in terms ofefficacy, tolerability and safety will be conducted.

(3) Pharmacogenetic (PGx) analyses may be performed on any bio-samplefrom subjects who have consented for PGx sampling. Subjectconfidentiality will be maintained. This analysis is optional.

Statistical Methods

Treatment population definitions:

Enrolled: All subjects who sign the informed consent form (ICF).

Intent-to-treat: All subjects randomly assigned to study drug. Subjectswill be analyzed according to the intervention to which they have beenrandomly assigned.

Safety All subjects randomly assigned to study drug and who take atleast 1 dose of study drug. Subjects will be analyzed according to theintervention they actually received

Pharmacokinetic (PK) Population All randomized subjects who receive anyamount of study drug and who have at least one post-baseline PK samplewill comprise the PK Population. Data will be analyzed based ontreatment received

Safety and Tolerability:

Continuous safety data are summarised with descriptive statistics(arithmetic mean, standard deviation [SD], median, minimum, and maximum)by treatment group. Categorical safety data are summarised withfrequency counts and percentages by treatment group. Adverse events arecoded using the most current Medical Dictionary for RegulatoryActivities (MedDRA) version available. The number of participantsexperiencing treatment-emergent adverse events as well as maximumseverity and relationship to study drug are summarized.

Laboratory evaluations, vital signs assessments, and ECG parameters aresummarised by treatment group and protocol-specified collection timepoint. A summary of change-from-baseline at each protocol-specified timepoint by treatment group are also presented. Concomitant medications arelisted by participant and coded using the most current World HealthOrganization drug dictionary. Medical history is listed by subject.Further details regarding presentation and analysis of safety data isdetailed in the Statistical Analysis Plan (SAP).

Pharmacokinetics:

Individual steady state atrasentan concentration data is listed andsummarised with descriptive statistics (sample size, arithmetic mean,SD, median, minimum, maximum, geometric mean, and geometric coefficientof variation). These data may be fed into a larger population PK modelof atrasentan. Correlations between atrasentan drug levels and safety orefficacy responses may be explored. Details regarding the statisticalanalyses of pharmacokinetic data are provided in the SAP.

Efficacy Evaluations:

Descriptive statistics are used to evaluate differences in demographicand baseline characteristics.

The primary efficacy endpoint is the change in proteinuria (UPCR) basedon 24-hour urine collection) from baseline to Week 24. The primaryanalysis will be conducted after approximately 270 subjects in theintent-to-treat (ITT) analysis set complete the Week 24 visit ordiscontinue from the study. The primary analysis will be tested based ona 2-sided significance level of 0.01. Baseline and Week 24 UPCR valueswill be estimated as the mean of the natural log of two separate samplescollected within 14 days apart prior to any study drug values.

The primary endpoint will be analyzed using a mixed-effects modelrepeated-measures (MMRM) model. The MMRM model will include change frombaseline of natural log UPCR at each postbaseline measurement asoutcomes. The model will also include the fixed effects of treatment,visit, and treatment-by-visit interaction, with covariates of baselinenatural log UPCR and baseline eGFR as continuous variables and regionrandomization stratification factors (region: Asia vs all otherregions). The covariance structure is assumed to be unstructured.Missing data will be assumed to be missing at random. UPCR datacollected after dialysis, use of SGLT-2 inhibitor, or use of prohibitedsystemic corticosteroids will not be included in the analysis. Theresulting analysis at 24-weeks will be used to assess efficacy.

Supportive and sensitivity analyses of the primary endpoint will beconducted to assess the robustness of the data and to evaluate theimpact of missing data. A supportive analysis will be conducted usingthe analysis of covariance (ANCOVA) methodology on the point estimate ofthe change from baseline to Week 24 in natural log UPCR. Missing datawill not be imputed and therefore assumed to be missing at random.Sensitivity analyses will be conducted including all on- andoff-treatment data through the 24-week endpoint analysis with an MMRMmodel as described in the primary analysis. Additionally, a tippingpoint analysis will be conducted to assess the impact of missing data onthe final endpoint. These sensitivity analyses will be further describedin the SAP. Subjects missing baseline UPCR will be excluded fromefficacy analyses. Subgroup analyses for clinically relevant demographicand baseline disease characteristics may be performed as specified inthe SAP.

Example 6. A Randomized, Double-Blind, Placebo-Controlled Study ofAtrasentan in Patients with IgA Nephropathy at Risk of Progressive Lossof Renal Function Study Overview

This study is a Phase 3, double-blind, placebo-controlled study tocompare the efficacy and safety of atrasentan, a selective endothelin-Areceptor antagonist, to placebo in patients with IgA nephropathy at riskof progressive loss of renal function. Approximately 320 patients willbe randomized 1:1 to receive atrasentan 0.75 mg daily or a matchedplacebo for 132 weeks while on a maximally tolerated and stable dose ofa RAS inhibitor (physician choice). Patients who are unable to tolerateRAS inhibition (up to 5% of study population) may be included Study armswill be stratified according to region, and baseline urinary proteinlevels.

Patients will have a telephone contact for safety evaluation at week 1and then return to clinic at week 2, 4, 6, 12, and then every 12 weeksthrough the completion of treatment at week 132 for safety and efficacyassessments. Pregnancy testing and patient counseling on contraceptionrequirements will occur monthly throughout the study for women ofchild-bearing potential. A negative pregnancy test result will beverified by a health care provider prior to the patient being approvedfor continued dosing. A positive or indeterminate pregnancy test resultwill lead to immediate hold of study medication and a repeat serum testat the clinical trial site within 2 days. Any positive serum pregnancytest will lead to permanent treatment discontinuation. In certainregions, with approval of the lead investigator and arrangement with thesponsor, patients may have an option for monthly urine pregnancy testingat a local point of care facility/lab or at-home urine pregnancy testingwith a virtual site-visit enabled by video/audio.

Patients that discontinue treatment prematurely before week 132 shouldremain on study for quarterly efficacy assessments to provide data forthe study endpoints. Patients will return to the clinic approximately 4weeks after the last dose of study drug. An additional blood draw shouldbe collected approximately 2 weeks after last dose of study drug. Thefinal study visit will occur at approximately week 136. Patientscompleting the study through Week 136 may be eligible to enroll in anextension study to receive open-label treatment with atrasentan under aseparate protocol.

Objectives:

The primary objective of this study is:

(1) To evaluate the effect of atrasentan versus placebo on proteinurialevels at week 24 compared to placebo.

The secondary objectives are:

(2) To evaluate the effect of atrasentan versus placebo on change frombaseline to week 136 (4 weeks post cessation of randomized treatment) inestimated glomerular filtration rate (eGFR);

(3) To compare two-year on-treatment rates of change in eGFR betweenatrasentan and placebo (eGFR slope week 12 to 120 of randomizedtreatment); and

(4) To compare the total on study rates of change in eGFR betweenatrasentan and placebo (eGFR slope from baseline to week 136).

The exploratory objectives are:

(5) To compare the percentage of patients achieving a proteinuriareduction of at least 50% from baseline and to a value less than 1 g/dayat week 24 between atrasentan and placebo;

(6) To evaluate the percentage of patients achieving a proteinuria levelof less than 0.3 g/day at week 24;

(7) To evaluate the percentage of patients experiencing at least a 30%reduction in eGFR during the study;

(8) To evaluate the percentage of patients experiencing at least a 40%reduction in eGFR during the study;

(9) To evaluate quality of life (QOL) of patients receiving atrasentancompared to placebo; and

(10) To evaluate the steady-state pharmacokinetics of atrasentan thatsupport exploratory exposure-response analyses.

Number of Subjects

Approximately 320 patients will be enrolled. Patients who discontinuefor reasons other than safety may be replaced at the discretion of thesponsor to ensure adequate data review after at least 6 weeks oftreatment.

Number of Study Sites: Approximately 120 sites worldwide

Criteria for Inclusion:

The following are the inclusion criteria. Patients must meet ALL of thefollowing inclusion criteria to be enrolled:

(1) male and female patients aged 18 and older;

(2) biopsy-proven diagnosis of IgA nephropathy not due to secondarycauses;

(3) receiving a maximally tolerated stable dose of a RAS inhibitor forat least 12 weeks prior to screening;

(4) Urine protein to creatinine (UPCR) of at least 1 g/g at screening.

(5) eGFR of at least 30 mL/min/1.73 m² at screening;

(6) willing to abide with highly effective forms of contraception asspecified in the protocol throughout the study; and

(7) willing and able to provide written informed consent and comply withall study visits and study procedures.

Exclusion Criteria

The following are the exclusion criteria; patients must meet NONE of thefollowing exclusion criteria to be enrolled:

(1) concurrent diagnosis of another cause of chronic kidney diseaseincluding diabetic kidney disease, hypertensive kidney disease oranother primary glomerulopathy. A history of well controlledhypertension is acceptable;

(2) presence of cellular glomerular crescents in >25% of glomeruli onkidney biopsy (if biopsy available within 6 months of screening) orclinical suspicion of rapidly progressive glomerulonephritis (RPGN);

(3) history of organ transplantation;

(4) use of systemic immunosuppressant medications (or investigationalagents) for at least 2 weeks in the past 6 months;

(5) blood pressure of >160 systolic or >100 diastolic at screening;

(6) history of heart failure or prior hospital admissions for conditionsrelating to fluid overload such as uncontrolled peripheral edema,pleural effusion, or ascites;

(7) history of clinically significant liver disease and transaminase orbilirubin values more than twice the upper limit of normal;

(8) hemoglobin below 9 g/dL at screening or prior history of bloodtransfusion for anemia within 3 months of screening;

(9) history of malignancy unless cancer free for at least 5 years ornonmelanoma skin cancer not requiring ongoing treatment;

(10) pregnancy, breast feeding, or intent to become pregnant during thestudy period for females;

(11) intent to father a child during the study period for males;

(12) have received any investigational or biologic agent within 1 month(or 5 half-lives of the agent, whichever is longer) prior to screening.If the investigational agent is a cytotoxic or immunosuppressive agentthen this washout period is 6 months;

(13) concurrent clinically significant, unstable, or uncontrolledcardiovascular, pulmonary, hepatic, renal, gastrointestinal,genitourinary, haematological, coagulation, immunological,endocrine/metabolic, or other medical disorder that, in the opinion ofthe Investigator, might confound the results of the study or poseadditional risk to the patient by their participation in the study;

(14) history of an alcohol or illicit drug-related disorder;

(15) Ongoing diagnosis of nephrotic syndrome with serum albumin <3 g/dLat screening; or

(16) Brain natriuretic peptide (BNP) value of >200 pg/mL at screening.

Note: Screening of a previously ineligible patient may be repeated withmedical monitor approval.

Test Product(s), Dose, and Mode of Administration:

Patients receive either: atrasentan 0.75 mg once daily by oraladministration; or matching placebo once daily by oral administration.

Atrasentan or placebo tablets will be supplied in a blinded manner inbottles with a 4-week supply (35 tablets to accommodate visit window).Patients will be instructed to take one tablet daily at approximatelythe same time, preferably in the morning. On days with study visits,patients should hold dosing until in clinic to enable pharmacokineticsampling prior to dose administration as applicable. Medication can betaken with or without food. No study drug dose reductions are permittedduring the study. Study drug should be temporarily halted or permanentlydiscontinued in the event of unacceptable toxicity that cannot bemanaged through supportive measures.

Treatment assignment will be determined by an interactive voice/webresponse system (IXRS). Patients, investigators, and the sponsor willnot have access to the assigned study treatment. In the event of amedical emergency the investigator will be able to receive the treatmentassignment through the IXRS system if required to provide optimal careof the patient. Randomization will be stratified by region (Asia vs allother regions), baseline urinary protein levels at screening (≥2 g/dayvs<2 g/day) and baseline eGFR (≥60 mL/min/1.73 m² vs<60 mL/min/1.73 m²).All patients will continue to receive a maximally tolerated and stabledose of RAS inhibitor (angiotensin converting enzyme inhibitor orangiotensin-receptor blocker) of the physician's choice as per standardof care. All efforts should be made to maintain a stable dosing regimenthroughout the study.

Duration of Treatment:

The study duration for an individual patient is approximately 2 yearsand 7 months (136 weeks) to include up to 4 weeks for screening, 132weeks of blinded study treatment, and 4 weeks for off-treatmentfollow-up. Patients may provide additional consent for pre-screening tooccur up to 6 months prior to the anticipated baseline visit; however,pre-screening assessments will have to be repeated for screening ifperformed outside of the 28-day screening window. The total duration ofthe study is expected to be 4-5 years. Enrolment is anticipated to last2 to 2.5 years with the last patient completing treatment and safetyfollow-up approximately 2 years and 7 months later.

Criteria for Evaluation: Efficacy

The primary efficacy endpoint is the change in proteinuria (urineprotein/creatinine ratio based on 24-hour urine collection) frombaseline to week 24 in atrasentan-treated patients as compared toplacebo-treated patients.

The key secondary efficacy endpoint is the rate of change in eGFR usingthe CKD-EPI creatinine equation from values at baseline through to thestudy completion visit (week 136) in atrasentan-treated patients ascompared to placebo-treated patients. Additional secondary efficacyendpoints include: rate of change in eGFR during two years on treatmentcalculated from values at baseline to week 136 in atrasentan-treatedpatients as compared to placebo-treated patients.

Exploratory endpoints include the percent of patients achievingproteinuria reduction to less than 1 g/day at week 24 and 50% decreasefrom baseline, percent of patients achieving proteinuria reduction toless than 0.3 g/day at week 24, percent of patients experiencing atleast a 30% reduction in eGFR during the study, and percent of patientsexperiencing at least a 40% reduction in eGFR during the study.

Criteria for Evaluation: Safety

Safety endpoints include:

(1) Type, incidence, severity, grading, seriousness, and relatedness ofadverse events;

(2) Incidence, severity, seriousness, and relatedness of adverse eventsof special interest including events of fluid overload; and

(3) Clinically significant changes in safety labs, ECGs, or physicalexamination findings (including vital signs).

Stopping Criteria

An independent data monitoring committee (DMC) will be appointed tomonitor the study. The DMC will meet periodically to review the safetyand will make recommendations regarding continuation, modification,suspension or termination of the study.

Patients will be required to discontinue study drug for the followingreasons:

(1) Evidence of pregnancy or noncompliance with protocol-specifiedcontraception or pregnancy monitoring;

(2) Initiation of high-dose steroids or other immunosuppressants orcytotoxic agents for treatment of IgAN; or

(3) Chronic dialysis for ESRD.

Other study drug stopping criteria may be based on investigator andpatient discretion that may include fluid overload syndromesuncontrolled with diuretics and medical management, and any suspectedstudy drug-related adverse event that represents unacceptable toxicity.All patients who stop study drug prior to week 132 will have blood draw2 weeks after discontinuation and a safety follow up visit after 4weeks. Patients should continue quarterly visits through week 136 forefficacy assessments only.

Statistical Methods Treatment Population Definitions:

Safety Population: All randomised participants who receive any amount ofstudy drug comprise the Safety Population for safety and exposureanalyses. Data will be analysed based on treatment received.

Intent-to-Treat: All randomised participants comprise theIntent-to-Treat Population for efficacy analyses. Data will be analysedbased on randomized treatment assignment.

PK Population: All randomised participants who receive any amount ofstudy drug and who have at least one post-baseline PK sample willcomprise the PK Population. Data will be analysed based on treatmentreceived.

Safety and Tolerability:

Continuous safety data are summarised with descriptive statistics(arithmetic mean, standard deviation [SD], median, minimum, and maximum)by treatment group. Categorical safety data are summarised withfrequency counts and percentages by treatment group. Adverse events arecoded using the most current Medical Dictionary for RegulatoryActivities (MedDRA) version available. The number of participantsexperiencing treatment-emergent adverse events as well as maximumseverity and relationship to study drug are summarized.

Laboratory evaluations, vital signs assessments, and ECG parameters aresummarised by treatment group and protocol-specified collection timepoint. A summary of change-from-baseline at each protocol-specified timepoint by treatment group are also presented. Concomitant medications arelisted by participant and coded using the most current World HealthOrganization drug dictionary. Medical history is listed by subject.Further details regarding presentation and analysis of safety data isdetailed in the Statistical Analysis Plan (SAP).

Pharmacokinetics:

Individual steady state atrasentan concentration data is listed andsummarised with descriptive statistics (sample size, arithmetic mean,SD, median, minimum, maximum, geometric mean, and geometric coefficientof variation). These data may be fed into a larger population PK modelof atrasentan. Correlations between atrasentan drug levels and safety orefficacy responses may be explored.

Efficacy Evaluations:

The primary and key secondary efficacy endpoints of the trial will beordered hierarchically and tested sequentially with gatekeepingprocedures. The primary endpoint will be tested first, acting as thegatekeeper, and if the null hypothesis is rejected at the 1%significance level (alpha=0.01), the key secondary endpoint will betested. The study will be deemed positive if the key secondary endpointnull hypothesis is rejected at the 5% significance level (alpha=0.05).

The primary efficacy endpoint is the change from baseline to eachpostbaseline visit up to week 24 in urinary protein to creatinine ratio(UPCR) as determined from 24-hour urine collection samples. As theprimary analysis is considering longitudinal results, the analysismethodology will be a mixed-effects model repeated-measures (MMRM)analysis of change from baseline to each postbaseline measurement of logUPCR. The model will include the fixed effects of treatment, visit, andtreatment-by-visit interaction, with covariates including randomizationstratification factors (region: Asia vs all other regions; baselineurinary protein levels at screening: ≥2 g/day vs<2 g/day, and baselineeGFR ≥60 mL/min/1.73 m² vs<60 mL/min/1.73 m²). The covariance structureis assumed to be unstructured. The resulting analysis at 24-weeks willbe used to assess efficacy. A supportive analysis will be conductedusing the analysis of covariance (ANCOVA) methodology on the pointestimate of the change from baseline to Week 24 in log UPCR. Thisanalysis will be performed on the primary endpoint to evaluate therobustness of the data. Missing data will not be imputed and thereforeassumed to be missing at random. Patients missing baseline UPCR will beexcluded from efficacy analyses. Further sensitivity analyses may beperformed to assess the impact of missing data. Subgroup analyses forclinically relevant demographic and baseline disease characteristicswill be performed as specified in the statistical analysis plan.

Following the collection and analysis of primary endpoint data at week24, patients will continue to receive their assigned blinded studytreatment up until week 132 and be followed off of study drug for anadditional 4 weeks to study completion at week 136 to collect additionaldata on eGFR decline.

The baseline and final eGFR values will be estimated as the mean of twosamples at each time point to reduce variability. The baseline referenceeGFR value will be determined from the mean of the screening andbaseline (prior to any study drug) values and the final eGFR value willbe estimated as the mean of two separate samples taken between 7 daysand 35 days (inclusive of visit windows) after last dose of study drug.Estimated GFR will be determined using the Chronic Kidney DiseaseEpidemiology Collaboration equation. The change from baseline in eGFRfor each patient will be determined by the absolute difference in valuesaveraged at baseline and Week 136, and then normalized for individualtime on study to decrease risk of bias from any differences betweengroups in the incidence of timing of early discontinuations from study.The endpoint will be analyzed by means of an ANCOVA with factors such astreatment group and randomization stratification factors included asindependent variables.

Example 7. A Phase 3, Randomized Placebo-Controlled Study of Atrasentanand Dapagliflozin in Patients with IgA Nephropathy

This example describes a Phase 3, Randomized, Double-Blind,Placebo-Controlled Study of Atrasentan in patients with IgA nephropathy)at risk of progressive loss of renal function despite treatment with amaximally tolerated and stable dose of a RAS inhibitor.

Study Overview

This example describes a randomized placebo-controlled efficacy andsafety study of atrasentan, a selective endothelin-A receptorantagonist, in combination with dapagliflozin, a sodium-glucose linkedtransporter 2 (SGLT-2) inhibitor, in patients with IgA nephropathy.Approximately 36 patients will be randomized 1:1:1 to receive atrasentan0.75 mg daily, atrasentan 0.75 mg daily and dapagliflozin 10 mg daily,or a matched placebo (for both atrasentan and dapagliflozin) for 4 weeks(plus a two week washout period), in addition to a maximally tolerateddose of a RAS inhibitor.

Patients will have a telephone contact for safety evaluation at week 1and will return to clinic at week 3 and at the end of the washout period(week 6) for safety and efficacy assessments.

Objectives:

The primary objective of this study is:

(1) To evaluate the effect of atrasentan in combination withdapagliflozin versus atrasentan alone on proteinuria levels compared toplacebo.

The secondary objectives are:

(2) To evaluate the effect on change from baseline in albumin tocreatinine ratio over the active treatment period and washout period;

(3) To evaluate the effect on change from baseline in body weight overthe active treatment period and washout period;

(4) To evaluate the effect on change from baseline in extracellularfluid (for example, measured by bio-impedance spectroscopy (Impedimed))over the active treatment period and washout period;

(5) To evaluate the effect on change from baseline in brain natriureticpeptide (BNP) levels over the active treatment period and washoutperiod; and

(6) To evaluate the effect on change from baseline in systolic bloodpressure over the active treatment period and washout period.

The exploratory objectives are:

(7) To compare the change in haematocrit from baseline to end of activetreatment;

(8) To compare the change in eGFR from baseline to end of activetreatment; and

(9) To compare the change in eGFR from baseline to end of wash outperiod (2 weeks after last dose of study drug).

Number of Subjects

Approximately 36 patients will be enrolled. Patients who discontinue forreasons other than safety may be replaced at the discretion of thesponsor to ensure adequate data review after at least 6 weeks oftreatment.

Number of Study Sites: Approximately 6 sites worldwide

Criteria for Inclusion:

The following are the inclusion criteria. Patients must meet ALL of thefollowing inclusion criteria to be enrolled:

(1) male and female patients aged 18 and older;

(2) biopsy-proven diagnosis of IgA nephropathy not due to secondarycauses;

(3) receiving a maximally tolerated stable dose of a RAS inhibitor forat least 4 weeks prior to screening;

(4) eGFR of at least 30 mL/min/1.73 m² at screening;

(5) willing to abide with highly effective forms of contraception asspecified in the protocol throughout the study; and

(6) willing and able to provide written informed consent and comply withall study visits and study procedures.

Exclusion Criteria

The following are the exclusion criteria; patients must meet NONE of thefollowing exclusion criteria to be enrolled:

(1) concurrent diagnosis of another cause of chronic kidney diseaseincluding diabetic kidney disease, hypertensive kidney disease oranother primary glomerulopathy. A history of well controlledhypertension is acceptable;

(2) concurrent diagnosis of type 1 diabetes or type 2 diabetes;

(3) brain natriuretic peptide (BNP) value of >200 pg/mL at screening;

(4) cardiovascular event within 3 months prior to screening;

(5) history of heart failure or prior hospital admissions for conditionsrelating to fluid overload such as uncontrolled peripheral edema,pleural effusion, or ascites;

(6) use of systemic immunosuppressant medications (or investigationalagents) for at least 2 weeks in the past 6 months;

(7) blood pressure of >160 systolic or >100 diastolic at screening;

(8) history of clinically significant liver disease and transaminase orbilirubin values more than twice the upper limit of normal;

(9) hemoglobin below 9 g/dL at screening or prior history of bloodtransfusion for anemia within 3 months of screening;

(10) history of malignancy unless cancer free for at least 5 years ornonmelanoma skin cancer not requiring ongoing treatment;

(11) have received any investigational or biologic agent within 1 month(or 5 half-lives of the agent, whichever is longer) prior to screening.If the investigational agent is a cytotoxic or immunosuppressive agentthen this washout period is 6 months;

(12) concurrent clinically significant, unstable, or uncontrolledcardiovascular, pulmonary, hepatic, renal, gastrointestinal,genitourinary, haematological, coagulation, immunological,endocrine/metabolic, or other medical disorder that, in the opinion ofthe Investigator, might confound the results of the study or poseadditional risk to the patient by their participation in the study;

(13) history of an alcohol or illicit drug-related disorder; or

(14) Ongoing diagnosis of nephrotic syndrome with serum albumin <3 g/dLat screening.

Note: Screening of a previously ineligible patient may be repeated withmedical monitor approval.

Test Product(s), Dose, and Mode of Administration:

Patients receive either: atrasentan 0.75 mg once daily by oraladministration; or atrasentan 0.75 mg and dapagliflozin 10 mg, each oncedaily by oral administration; or matching placebo once daily by oraladministration.

Treatment assignment will be determined by an interactive voice/webresponse system (IXRS). Patients, investigators, and the sponsor willnot have access to the assigned study treatment. In the event of amedical emergency the investigator will be able to receive the treatmentassignment through the IXRS system if required to provide optimal careof the patient. Randomization will be stratified by region (Asia vs allother regions), baseline urinary protein levels at screening (≥2 g/dayvs<2 g/day) and baseline eGFR (≥60 mL/min/1.73 m² vs<60 mL/min/1.73 m²).All patients will continue to receive a maximally tolerated and stabledose of RAS inhibitor (angiotensin converting enzyme inhibitor orangiotensin-receptor blocker) of the physician's choice as per standardof care. All efforts should be made to maintain a stable dosing regimenthroughout the study.

Criteria for Evaluation: Efficacy

The primary efficacy endpoint is the change in proteinuria (urineprotein/creatinine ratio based on 24-hour urine collection) frombaseline to week 4 in atrasentan-treated patients and inatrasentan+dapagliflozin treated patients as compared to placebo-treatedpatients.

Criteria for Evaluation: Safety

Safety endpoints include:

(1) Type, incidence, severity, grading, seriousness, and relatedness ofadverse events;

(2) Incidence, severity, seriousness, and relatedness of adverse eventsof special interest including events of fluid overload; and

(3) Clinically significant changes in safety labs, ECGs, or physicalexamination findings (including vital signs).

Stopping Criteria

An independent data monitoring committee (DMC) will be appointed tomonitor the study. The DMC will meet periodically to review the safetyand will make recommendations regarding continuation, modification,suspension or termination of the study.

Safety and Tolerability:

Continuous safety data are summarised with descriptive statistics(arithmetic mean, standard deviation [SD], median, minimum, and maximum)by treatment group. Categorical safety data are summarised withfrequency counts and percentages by treatment group. Adverse events arecoded using the most current Medical Dictionary for RegulatoryActivities (MedDRA) version available. The number of participantsexperiencing treatment-emergent adverse events as well as maximumseverity and relationship to study drug are summarized.

Laboratory evaluations, vital signs assessments, and ECG parameters aresummarised by treatment group and protocol-specified collection timepoint. A summary of change-from-baseline at each protocol-specified timepoint by treatment group are also presented. Concomitant medications arelisted by participant and coded using the most current World HealthOrganization drug dictionary. Medical history is listed by subject.

Efficacy Evaluations:

The primary efficacy endpoint is the change from baseline to end ofstudy proteinuria levels as determined from 24-hour urine collectionsamples. Following the collection and analysis of primary endpoint dataat week 4, patients will be followed off of study drug for an additional2 weeks (the washout period) to collect additional data on proteinurialevels.

Example 8. In Vivo Study Using a Mouse Model of IgA Nephropathy

The grouped ddY (g-ddY) mouse is a spontaneous model of early-onset IgAnephropathy characterized by IgA immune complex deposition in themesangium of the kidney, leading to significant proteinuria, glomerularhypercellularity, mesangioproliferative glomerular lesions, glomerularsclerosis and reduced kidney function, all hallmarks of human IgAnephropathy (J Am Soc Nephrol 23: 1364-1374, 2012). Consistent withhuman IgA nephropathy, the mesangial IgA deposits in the g-ddY mouse arealso accompanied by deposition of IgG and complement C3.

Methods

Approximately 6-week-old g-ddY mice were administered varying doses ofatrasentan hydrochloride (10, 20, or 30 mg/kg/day, n=3/group) indrinking water for a duration of approximately 5 days. A separate groupof 6-week-old male control g-ddY mice (n=2) received regular drinkingwater without atrasentan. Proteinuria was assessed as urine albumin tocreatinine ratio (UACR), at baseline prior to atrasentan administrationand on the final day of atrasentan administration. The effect ofatrasentan on UACR was compared relative to baseline, within anindividual mouse, and also to the control group who did not receiveatrasentan.

RNA-sequencing (RNA-seq) was used to assess changes in gene expressionin kidney tissue following administration of atrasentan to g-ddY mice at0 (control), 10, 20, or 30 mg/kg/day. RNA isolation and sequencinglibraries were generated using standard protocols. Sequencing wasperformed using an Illumina NextSeq500 using manufacturer's instructionswith a read length of 75 bp and a read depth of 30 million. Raw data wasassembled to FASTQ files. Following QC analysis, high quality sequencingreads (e.g., reads with a Phred quality score >36) were aligned to themouse reference genome (GRCm38-mm10) using the default parameters of thealign algorithm and transcript count tables of each library wasgenerated using the default parameters of the featureCounts algorithm ofthe RSubread package (Liao Y, Smyth G K, Shi W, 2019). High qualitysequencing reads mapped to 27,129 genes. Following additional QC of lowcount filtering (i.e. transcripts <10 counts across all samples), 16,207genes were identified and used for further analysis. The remainingsequencing read counts of the included 16,207 genes were normalized tolibrary size, log-transformed, and were quantified as counts per million(CPM). Select RNA-seq counts were validated using qPCR.

Sequencing read counts (CPM) were used for identification ofdifferential gene expression between specified populations using thequasi-likelihood methodology of the edgeR package. Genes were considereddifferentially expressed between populations if they had an adjustedp-value (FDR of less than 0.05). MA plots were generated using edgeR.Heatmaps and other plots were generated using the gplots and ggplot2package.

Analysis of gene pathways was performed using Qiagen's Ingenuity PathwayAnalysis (IPA) and Gene Set Enrichment Analysis (Subramanian, et al.,Gene set enrichment analysis: A knowledge-based approach forinterpreting genome-wide expression profiles. Proc. Nat. Acad. Sci. USA,102(43), 15545-15550 (2005). The outcome from the differential geneexpression analysis was uploaded into the IPA system for core analysisand then overlaid with the global molecular network in the Ingenuitypathway knowledge base (IPKB). IPA was performed to identify canonicalpathways and gene networks that are most significant to thedifferentially expressed genes identified using the analysis above. Geneexpression data input into IPA did not include a fold-change thresholdand the P-value filter was set at <0.001. The resulting enriched genepathways were considered significant with Benjamin-Hochberg adjustedp-value <0.001. IPA analysis output was summarized in a heatmap withactivation z-scores representing pathway enrichment in differentpopulations. See FIG. 6. For GSEA, the differentially expressed genelist from edgeR (Robinson et al., A Bioconductor package fordifferential expression analysis of digital gene expression data.Bioinformatics, 26(1), 139-140 (2009) was rank ordered based on theF-statistic without arbitrary logFC or p-value thresholds. GSEA wasimplemented using the fgsea package (Korotkevich, et al., BioRxiv 2016,Fast gene set enrichment analysis. 1-29. https://doi.org/10.1101/060012)and the Hallmark gene set collection of MSigDB (Liberzon et al., TheMolecular Signatures Database Hallmark Gene Set Collection. CellSystems, 1(6), 417-425 (2015)).

Results

Healthy Balb/c mice without IgA nephropathy (the appropriate controlstrain for the g-ddY mouse) have very low, nearly undetectable levels ofUACR, even through 2 years of age (Tsaih et al., 2010, KidneyInternational). At baseline, the grouped g-ddY mice used in this studyhad substantial proteinuria, with a mean (±standard error of the mean(SEM)) UACR level of 293.1±16 mg/g. The baseline albuminuria was wellmatched across treatment groups, with no significant differences betweenany groups (FIG. 4).

Effect of Atrasentan on UACR: Comparison to Baseline

Atrasentan treatment for approximately 5 days, at 30 mg/kg/daysignificantly (P=0.0002, paired two-sided t-test) reduced albuminuriafrom baseline (FIG. 4). Albuminuria was reduced from baseline to asimilar extent by treatment with atrasentan at 20 mg/kg/day, howeverthis effect just failed to reach statistical significance (P=0.0593)(FIG. 4). Albuminuria was not significantly changed from baseline incontrol g-ddY mice who did not receive atrasentan, or in the group ofmice treated with atrasentan at 10 mg/kg/day; although albuminuria wasreduced in ⅔ mice in this 10 mg/kg/day group (FIG. 4).

Effect of Atrasentan on UACR: Comparison to Control

The change in albuminuria from baseline (percent (%) change frombaseline), following approximately 5 days of treatment with atrasentanor control is shown in FIG. 5. Atrasentan reduced UACR from baseline by28±44%, 62±8% and 63±6% at 10, 20 and 30 mg/kg/day, respectively. Theeffect of atrasentan to reduce UACR from baseline at 20 mg/kg/day(P=0.0498, un-paired t-test) and 30 mg/kg/day (P=0.029), wasstatistically significant compared to the control group (0 mg/kg/day)(FIG. 5). However, the effect of atrasentan at 10 mg/kg/day was notsignificant, although again proteinuria was reduced in ⅔ mice in this 10mg/kg/day group (FIG. 5).

Gene Expression Analysis Following Treatment with Atrasentan

Analysis of RNA-seq data provided the top 100 differentially expressedgenes in kidney tissue following administration of atrasentan to g-ddYmice at 0 (control), 10, 20, or 30 mg/kg/day clustered according todisease and atrasentan treatment condition, as shown in FIG. 6 and Table5. Pairwise comparison between 10 mg/kg/day versus control, 20 mg/kg/dayversus control, and 30 mg/kg/day versus control showed dose-dependentgene expression changes (FIGS. 7A-7C). FIG. 7A shows differentiallyexpressed genes for 10 mg/kg/day versus control where 25 genes areupregulated (+log-fold-change) and 15 genes are downregulated(−log-fold-change). Upregulated and downregulated genes from FIG. 7A areshown in Table 1. FIG. 7B shows differentially expressed genes for 20mg/kg/day versus control where 214 genes are upregulated(+log-fold-change) and 281 genes are downregulated (−log-fold-change).The top 25 upregulated and top 25 downregulated genes from FIG. 7B areshown in Table 2. FIG. 7C shows differentially expressed genes for 30mg/kg/day versus control where 910 genes are upregulated(+log-fold-change) and 768 genes are downregulated (−log-fold-change).The top 25 upregulated and top 25 down regulated genes from FIG. 7C areshown in Table 3.

Comparing gene expression changes in atrasentan treated mice with ahuman specific Atra/ET1 gene signature showed that 53 of 60 genesoverlapped. Specifically, FIGS. 8A-8C show differential gene expressiongenes (p-value <0.05) annotated as being part of the 60 genes associatedwith an ET1 gene signature and that are differentially expressed in micetreated with atrasentan. Gene expression analysis showed ET1 (End1) wasdownregulated across all three doses (FIGS. 8A-8C). In addition, Ednraand Ednrb were downregulated at 30 mg/kg/day. Of the genes thatoverlapped between mouse and human and were differentially expressedwhen comparing 10 mg/kg/day atrasentan treatment to untreated control,Ccnd1, Ppara, Agt, Map2k2 and Hoxa9 were upregulated while Grk5, Mllt3,Klf2, Pparg, Hgf, Edn1, Fos, Ptgs2, and IL-1b were downregulated (FIG.8A). Looking at the overlapping genes that were differentially expressedwhen comparing 20 mg/kg/day atrasentan treatment to untreated control,FIG. 8B shows Ccdn1, Cat, Ppara, Map2k2, Hoxa9, Agt, and Egfr wereupregulated while Nfatc1, Grk5, Klf2, Cav1, Pparg, Itga5, Myc, Ednra,Sell, Hgf, Mmp2, Edn1, Syk, Fos, and IL-1b were downregulated. Finally,differentially expressed genes that overlapped between species whencomparing 30 mg/kg/day atrasentan treatment to untreated controlsincluded upregulated genes Ccnd1, Agt, Cat, Map2k2, Hoxa9, and Ppara anddownregulated genes Ednrb, Myc, Mllt3, Hdac7, Ednra, Itga5, Grk5, Pparg,Klf2, Hgf, Mmp2, Sell, Sphk1, Fos, Syk, Edn1, and IL-1b (FIG. 8C).

Ingenuity pathway analysis (IPA) and Gene Set Enrichment Analysis (GSEA)were applied to systematically identify pathways associated with genesdifferentially expressed at different dosages of atrasentan. In IPA,canonical pathways significantly enriched that include an adjustedp-value less than 0.001 without a filter on the activation z-score areshown in Table 4. IPA analysis revealed that atrasentan treatment at 30mg/kg/day reduced NF-κB signaling, IL-6 pathway signaling (e.g., STAT3),and PDGF signaling. Tables 9-11 show specific signaling componentsimpacted by atrasentan treatment for each of NF-κB signaling (Table 9),IL-6 signaling (Table 10), and PDGF signaling (Table 11). In addition,IPA demonstrated that atrasentan treatment induced gene expressionchanges that work synergistically in pathways of rescue including, forexample, reduction in inflammatory and proliferative pathways. Thepositive and negative enriched gene sets from GSEA following atrasentantreatment are shown in Table 5. As shown in Table 5, GSEA identifiedconcordant negative enrichment (NES <−1.5 and adjusted p-value <0.05)indicating under-expression of hallmark signaling pathways includingTNFα signaling via NF-κB, epithelial mesenchymal transition,inflammatory response, IL6-JAK-STAT3 signaling, and gene sets associatedwith cell-proliferation (mitotic spindle and G2M checkpoint) followingtreatment of g-ddY mice with atrasentan at 10, 20, and 30 mg/kg/day.

Upstream regulator analysis using Ingenuity Pathway identified thepredicted activation/inactivation state of potential upstream regulatorymolecules in relation to the biological processes and pathways found tobe differentially regulated in the 30 mg/kg/day of atrasentan (Tables 6,7 and 8). Tables 6, 7 and 8 show the reduction in pro-inflammation,fibrosis, and cell proliferation was the result of predicted inhibitionof one or more positive regulators of inflammation, fibrosis, and cellproliferation and/or predicted activation of one or more negativeregulators of inflammation, fibrosis, and cell proliferation. Predictedinhibition and activation of cytokines, growth factors, transcriptionalregulators, ligand-dependent nuclear receptors, transmembrane receptors,and G-coupled protein receptors, can include, without limitation, EDN1,CCLS, CD40LG, CSF2, CSF3, CXCL2, IFNA2, IFNG, OSM, PF4, PRL, tumornecrosis family (TNF, TNFSF11, TNFSF12), TSLP, WNT3A, the interleukincytokine family (IL1A, IL1B, IL2, IL3, IL6, IL10, IL21, IL33), TGFB1,TGFB2, TGFB3, IGF1, HGF, AGT, EGF, NRG1, PDGFB, VEGFA, FGF23, BMP2,GDF2, FGF2, IL6R, EDNRA, toll-like receptor family (TLR3, TLR4, TLR9),JUNB, YAP1, TEAD1, TEAD2, TEAD3, TEAD4, NFKB1, JUNB, STAT1, STAT3,STAT4, SMAD2, SMAD3, and SMAD4.

As shown in Table 4, 5, and 8, treatment with 30 mg/kg/day of atrasentanreduced NF-κB signaling. Components of NF-κB signaling differentiallyexpressed following treatment with 30 mg/kg/day of atrasentan can befound in Table 9. Table 9 shows that the reduction in NK-κB signalingwas the result of a decrease of one or more positive regulators of NF-κBsignaling and/or increase of one or more negative regulators of NF-κBsignaling. Table 9 also shows that the reduction in NK-κB signaling wasalso the result of modulation of one or more core components of NF-κBsignaling such that NF-κB signaling was reduced. NF-κB signaling pathwaycomponents can include, without limitation, CHUK, FADD, IKBKB, IKBKG,IL1A, IL1R1, MAP3K1, MAP3K14, MAP3K7, MYD88, NFKB1, NFKBIA, RELA, RIPK1,TAB1, TNF, TNFAIP3, TNFRSF1A, TNFRSF1B, TRADD, and TRAF6 (see, e.g.,BIOCARTA NFKB_PATHWAY).

As shown in Tables 5 and 10, treatment with 30 mg/kg/day of atrasentanreduced IL-6 signaling. Components of IL-6 signaling differentiallyexpressed following treatment with 30 mg/kg/day of atrasentan can befound in Table 10. Table 10 shows the reduction in IL-6 signaling wasthe result of a decrease of one or more positive regulators of IL-6signaling and/or increase of one or more negative regulators of IL-6signaling. Table 10 also shows the reduction in IL-6 signaling was alsothe result of modulation of one or more core components of IL-6signaling such that IL-6 signaling was reduced. IL-6 signaling pathwaycomponents can include, without limitation, AGT, AKT1, BAD, BCL2L1,CREBBP, CRP, GAB1, GRB2, GSK3B, HCK, HDAC1, IL6, IL6R, IL6ST, IRF1,JAK1, JAK2, JUNB, MAP2K1, MAP2K2, MAP2K4, MAP3K7, MAPK1, MAPK3, NCOA1,NLK, NR2F6, PIK3R1, PIK3R2, PRDM1, PRKCD, PTPN11, RAC1, RPS6KB1, SHC1,SOCS3, SOS1, STAT1, STAT3, TIMP1, TYK2, VAV1, and VIP (see, e.g., WikiPathway IL6_SIGNALING_PATHWAY).

As shown in Tables 4 and 11, treatment with 30 mg/kg/day of atrasentanreduced PDGF signaling. Components of PDGF signaling differentiallyexpressed following treatment with 30 mg/kg/day of atrasentan can befound in Table 11. Table 11 shows the reduction in PDGF signaling wasthe result of a decrease of one or more positive regulators of PDGFsignaling and/or an increase of one or more negative regulators of PDGFsignaling. Table 11 also shows the reduction in PDGF signaling was alsothe result of modulation of one or more core components of PDGFsignaling such that PDGF signaling was reduced. PDGF signaling pathwaycomponents can include, without limitation, ARFIP2CDC42, CHUK, ELK1,FOS, GRB2, HRAS, JAK1, JUN, MAP2K1, MAP2K4, MAP3K1, MAPK1, MAPK3, MAPK8,MT-CO2, NFKB1, NFKBIA, PAK1, PDGFA, PDGFB, PDGFRB, PIK3R1, PLA2G4A,PLCG1, PTPN11, RAC1, RAF1, RASA1, RHOA, SHC1, SOS1, SRC, SRF, STAT1,STAT3, TIAM1, VAV1, VAV2, and WASL (see, e.g., Wiki PathwayPDGF_PATHWAY). For example, atrasentan can inhibit the expression and/oractivity of one or more PDGF signaling pathway components selected fromthe group of ARFIP2CDC42, CHUK, ELK1, FOS, GRB2, HRAS, JAK1, JUN,MAP2K1, MAP2K4, MAP3K1, MAPK1, MAPK3, MAPK8, MT-CO2, NFKB1, NFKBIA,PAK1, PDGFA, PDGFB, PDGFRA, PDGFRB, PIK3CG, PIK3R1, PLA2G4A, PLCG1,PTPN11, RAC1, RAF1, RASA1, RHOA, SHC1, SOS1, SRC, SRF, STAT1, STAT3,TIAM1, VAV1, VAV2, and WASL.

As shown in Tables 5 and 12, treatment with 30 mg/kg/day of atrasentanreduced cell proliferation associated signaling. Components of cellproliferation associated signaling (mitotic spindle and G2M checkpoint)differentially expressed following treatment with 30 mg/kg/day ofatrasentan can be found in Table 12. Table 12 shows the reduction incell proliferation associated signaling (mitotic spindle and G2Mcheckpoint) was the result of a decrease of one or more positiveregulators of cell proliferation associated signaling (mitotic spindleand G2M checkpoint) and/or an increase of one or more negativeregulators of cell proliferation associated signaling (mitotic spindleand G2M checkpoint). Table 12 also shows the reduction in cellproliferation associated signaling (mitotic spindle and G2M checkpoint)was also the result of modulation of one or more core components of cellproliferation associated signaling (mitotic spindle and G2M checkpoint)such that cell proliferation associated signaling (mitotic spindle andG2M checkpoint) was reduced. Cell proliferation associated signalingpathway (mitotic spindle and G2M checkpoint) components can include,without limitation, CCDC88A, SORBS2, RHOT2, EPB41L2, CEP192, BCAR1,PPP4R2, TUBA4A, OPHN1, CNTRL, TIAM1, NIN, MYH10, FLNA, RAPGEFS, FGD6,MYO1E, VCL, ITSN1, SMC3, MYH9, SLC12A2, DMD, CCND1, ARID4A, KMTSA, ATRX,SLC38A1, SLC7A1, YTHDC1, and MEIS1 (see, e.g., MSigDB HALLMARK G2MCHECKPOINT and HALLMARK MITOTIC SPINDLE).

As shown in Tables 5 and 13, treatment with 30 mg/kg/day of atrasentanreduced inflammatory response signaling. Components of inflammatoryresponse signaling differentially expressed following treatment with 30mg/kg/day of atrasentan can be found in Table 13. Table 13 shows thereduction in inflammatory response signaling was the result of adecrease of one or more positive regulators of cell proliferationassociated signaling inflammatory response signaling and/or an increaseof one or more negative regulators of inflammatory response signaling.Table 13 also shows the reduction in inflammatory response signaling wasalso the result of modulation of one or more core components ofinflammatory response signaling such that inflammatory responsesignaling was reduced. inflammatory response signaling pathwaycomponents can include, without limitation, RNF144B, ROS1, SLC7A2, F3,NFKBIA, HPN, SLC4A4, CHST2, IL1B, CDKN1A, BTG2, CSF1, SLC11A2, EIF2AK2,EDN1, NFKB1, SERPINE1, SLC7A1, CCL20, IL1R1, LY6E, and GABBR1 (see,e.g., MSigDB HALLMARK INFLAMMATORY RESPONSE).

Cross-validation of the impact of atrasentan treatment on hallmark genesets across two models (g-ddY mouse model of IgAN and ET-1 treated HRMC)is shown in FIG. 10. FIG. 10 illustrates the concordance in negativeenrichment scores indicating the under-expression of hallmark gene sets(i.e. TNFα signaling via NF-κB, mitotic spindle, inflammatory response,epithelial mesenchymal transition, IL2-STAT5 signaling, IL6-JAK-STAT3signaling, G2M checkpoint, E2F targets, complement, apoptosis, andallograft rejection) as a result of atrasentan treatment in the g-ddYand HRMC model systems. Enriched gene sets following the treatment ofHRMC with ET-1 compared to untreated HRMC (control) shows positiveenrichment scores indicating the over-expression of the same hallmarkgene sets (i.e. TNFα signaling via NF-κB, mitotic spindle, inflammatoryresponse, epithelial mesenchymal transition, IL2-STAT5 signaling,IL6-JAK-STAT3 signaling, G2M checkpoint, E2F targets, complement,apoptosis, and allograft rejection).

Analysis of mesangial cell associated gene signature is shown in FIG.11A and FIG. 11B. FIG. 11A shows the overlapping genes betweendifferentially expressed genes following treatment of g-ddY mice withatrasentan at 30 mg/kg/day compared to treatment of atrasentan at 0mg/kg/day (control), differentially expressed genes following treatmentof HRMC with atrasentan at 25 nM in the presence of ET-1, and apublished human mesangial single cell signature (Lake et al., 2018).FIG. 11B illustrates the increased and/or decreased expression levels of44 genes associated with mesangial cell signature in g-ddY mice treatedwith atrasentan at 0 mg/kg/day (control) and decreased and/or increasedexpression levels following treatment of g-ddY mice with atrasentan at10, 20, and 30 mg/kg/day. Components of the mesangial cell associatedgene signature is shown in Table 14.

Assessment of potential biomarkers based on the RNA-seq data is shown inFIGS. 12A-12B. FIG. 12A summarizes mouse versus human translatabilityamong the selected biomarkers: ALPL, AMN, ASL, CRYL1, DPEP1, GAPDH,IGF1, PDZK1IP1, PEBP1, SELENBP1, SLC19A1, and SLC6A19. FIG. 12A showswhich of the potential biomarkers were present in plasma/serum and/orurine and whether the potential biomarker was expressed in both mouseand human. As the RNA-seq data was generated in mouse, filteringpotential biomarkers based on a set of criteria, as seen in FIG. 12A,was used to limit potential future issue with lack of translatability.Gene expression changes (log 2(CPM)) for each potential biomarker wasplotted for control and atrasentan treatment at 10 mg/kg/day, 20mg/kg/day and 30 mg/kg/day (FIG. 12B). Based on the criteria in FIG. 12Aand the RNA-seq data in FIG. 12B, IGF1, DPEP1, ASL, AMN, ALPL andSLC6A19 were identified as potential biomarkers.

In summary, the gene expression analysis following atrasentan treatmentshowed treatment induced changes in genes that work synergistically inpathways of rescue (e.g., reduction in inflammatory and proliferativepathways). In addition, the RNA-seq data identified potential biomarkers(e.g., IGF1, DPEP1, ASL, AMN, ALPL, and SLC6A19) that can be usedalongside ET-1 (EDN1), TNF, and FGF-2. Note that the overalldose-dependent changes in gene expression following treatment suggestthat biomarkers and functional UACR readout can be used in conjunction.

Tables

TABLE 1 Top 40 DEGs (25 upregulated and 15 downregulated) in Atrasentan10 mg/kg/day vs. control. Top 25 Upregulated Genes Top 15 DownregulatedGenes Symbol logFC P Value FDR Symbol logFC PValue FDR Cryl1 0.5206467.06E−05 0.035738 Apoc1 −1.92878 0.000107 0.045268 Pdzk1ip1 0.5307546.24E−05 0.034124 Nr4a1 −1.52471 1.11E−05 0.02068 Slc7a9 0.5455799.00E−05 0.040481 Gem −1.50783 4.30E−05 0.028806 Bckdha 0.5491723.73E−05 0.028796 Gm15910 −1.39721 3.43E−05 0.027787 Gapdh 0.6256881.93E−05 0.021512 Cish −1.30133 2.79E−06 0.015098 Slc19a1 0.6450597.31E−05 0.035903 Dbp −1.20474 1.42E−05 0.020966 Dpep1 0.647598 4.23E−050.028806 Pfkfb3 −1.16606 5.04E−06 0.016337 Mnt 0.666881 2.82E−050.025402 Ccn1 −1.08749 6.74E−05 0.035252 Hacl1 0.680475 0.0001180.047742 Mmd −1.01248 1.02E−06 0.008642 Frmd4b 0.686272 5.76E−050.034124 Igf1 −0.87229 2.12E−05 0.021512 Mapt 0.703518 8.51E−05 0.040481Filip1l −0.81857 2.44E−05 0.023255 Asl 0.707013 1.15E−05 0.02068 Pde3b−0.66114 4.28E−05 0.028806 Bmp6 0.714133 1.99E−05 0.021512 Mef2c−0.62169 9.17E−05 0.040481 Pdzd3 0.726782 2.05E−05 0.021512 Zeb2−0.56382 0.000109 0.045268 Sirt3 0.737604 9.11E−06 0.02068 Ccn2 −0.537915.98E−05 0.034124 Slc5a2 0.747833 3.07E−05 0.026184 S100g 0.7546045.84E−05 0.034124 Slc6a19 0.85668 1.82E−05 0.021512 Slc34a3 0.892349.24E−05 0.040481 Acnat2 0.917366 4.44E−05 0.028806 Ethe1 0.9410257.89E−06 0.02068 Slc16a1 0.980825 1.07E−06 0.008642 Ndrg1 0.9887324.13E−06 0.016337 Amt1 1.039427 6.32E−05 0.034124 Npas2 1.41519 1.37E−050.020966

TABLE 2 Top 50 DEGs (25 upregulated and 25 downregulated) in Atrasentan20 mg/kg/day vs. control Top 25 Upregulated Genes Top 25 DownregulatedGenes Symbol logFC PValue FDR Symbol logFC PValue FDR Glis1 0.8775690.000497 0.033022 Lep −5.90482 0.000285 0.027287 Klf15 0.87985 9.74E−050.020853 Otop1 −5.80751 0.000198 0.026094 Ndrg1 0.894577 1.19E−050.011184 Paral1 −5.13251 0.000207 0.026429 Prodh 0.898674 5.01E−050.0167936 Sprr2a3 −4.40323 0.00025 0.026639 Lhx1 0.925842 0.0003820.029887 C5ar2 −4.27061 0.000429 0.031245 Pfn3 0.93398 0.000559 0.033736Bche −4.06216 0.000249 0.026639 Slc34a3 0.934522 5.82E−05 0.017159 Iv1−3.8887 0.000596 0.034028 Cndp1 0.937324 0.000184 0.025434 Dio2 −3.823285.45E−05 0.016936 Mom2 0.937472 0.000256 0.026639 Ebf2 −3.71451 0.0003160.028532 Tmem37 0.943864 8.88E−05 0.019978 Plekhs1 −3.66111 0.0001550.023432 Pvalb 0.944123 0.001322 0.047017 Apo16 −3.61598 2.86E−050.016936 Slc6a19 0.958468 5.59E−06 0.009287 Ccl20 −3.56331 0.0005670.033736 Aspdh 0.964605 0.000115 0.021249 Ankef1 −3.38241 0.0003470.029573 Hao2 0.985461 0.00074  0.037501 Ly6d −3.37673 0.000848 0.040407Cyp4b1-ps2 1.004361 0.000176 0.025048 Nat81 −3.26334 0.000359 0.029887Npas2 1.011763 0.000377 0.029887 Krt5 −3.04432 0.001278 0.046755 Clec2h1.077782 0.000267 0.026652 Il1b −3.02945 0.000263 0.026639 Dnase11.085844 0.000173 0.024747 A530016L24Rik −3.02722 0.000155 0.023432 Ftcd1.086278 0.000998 0.042127 Ccl8 −2.99918 0.000577 0.033797 Frs3 1.0937870.000444 0.031697 Abcd2 −2.68553 9.78E−06 0.011184 Reni 1.1174960.000566 0.033736 Lgals12 −2.53645 5.63E−05 0.016936 Fcarnr 1.1820321.22E−05 0.011184 Adcy2 −2.48445 0.000234 0.026429 1700101I11Rik 1.657750.000966 0.041578 Prkar2b −2.46803 0.000194 0.026008 Igfbp1 2.8200360.000497 0.033022 Cadm3 −2.43878 0.001079 0.04323 Hrg 4.888319 0.0001920.025954 Tns4 −2.38111 0.00111  0.043442

TABLE 3 Top 50 DEGs (25 upregulated and 25 downregulated) in Atrasentan30 mg/kg/day vs. control Top 25 Upregulated Genes Top 25 DownregulatedGenes Symbol logFC P Value FDR Symbol logFC PValue FDR Frs3 1.175840.000241 0.01131 Clec4d −4.65021 0.004497 0.046687 Slc16a1 1.1782851.26E−07 0.000796 Adamts17 −3.19106 0.001153 0.023207 Ndrg1 1.1972765.02E−07 0.000796 A530016L24Rik −3.0348 0.000191 0.010045 G6pc 1.2069710.000984 0.021884 Arhgap36 −2.90784 0.00497  0.04903 Ethe1 1.2105714.78E−07 0.000796 Rnf182 −2.81664 0.00456  0.046915 Calb1 1.2112361.55E−05 0.003454 Fam163b −2.78491 5.34E−05 0.00548 Clec2h 1.215149.25E−05 0.00704 Igkv3-5 −2.72187 0.004931 0.048844 Slc6a19 1.2382293.26E−07 0.000796 Cxcl2 −2.6664 0.004478 0.04664 Pgam2 1.24014 1.46E−050.003454 Xirp2 −2.61909 0.002983 0.037627 Mogat2 1.285346 0.0004440.015136 Ccl20 −2.47108 0.004346 0.045771 Nepn 1.290765 0.0005940.017346 Tns4 −2.44271 0.001081 0.022699 Nkx3-1 1.29376 0.0026910.035636 Kcnk2 −2.43099 0.001377 0.025337 S100g 1.295748 1.68E−070.000796 Plch2 −2.42276 0.003667 0.041761 Misp3 1.307023 0.0007550.019283 Ebf2 −2.38601 0.003955 0.043366 Cyp27b1 1.350092 0.00012 0.008122 E030018B13Rik −2.37324 5.79E−07 0.000796 Pvalb 1.3885864.38E−05 0.004946 Ankef1 −2.3582 0.003009 0.037799 Npas2 1.3935571.65E−05 0.003454 Adcy2 −2.32735 0.000415 0.014673 Chad1 1.4095040.003796 0.042633 Il1b −2.26303 0.001846 0.029185 Slc34a3 1.4359925.69E−07 0.000796 Stx1b −2.24452 0.000525 0.016224 Cyp2d26 1.4636250.00016 0.00927 Atf3 −2.21952 0.002193 0.031678 Reni 1.530684 4.64E−050.005079 Kif1a −2.10828 0.000147 0.008889 1700101I11Rik 1.6133760.00129  0.024488 1700003D09Rik −2.08702 0.003352 0.039918 Cyp2d37-ps2.922005 0.002459 0.033932 Gm20026 −2.0755 0.002092 0.031103 Igfbp13.447021 5.49E−05 0.005512 Wnt10a −2.07117 0.003982 0.043533 Bhmt4.173438 0.000315 0.012685 Fat3 −2.06711 0.000583 0.017174

TABLE 4 The enriched signaling pathways following treatment of g-ddYmice with atrasentan at 0 (control), 10, 20 or 30 mg/kg/day. Z-scorevalues indicate the magnitude of the effect seen at each dosage.Canonical Pathways Atra_10 mg Atra_20 mg Atra_30 mg OxidativePhosphorylation −0.309 −0.283 7.02 Folate Polyglutamylation −1 0.4471.342 PTEN Signaling −0.426 1.029 2.214 Apelin Cardiac FibroblastSignaling Pathway 1.134 0.577 1.732 NRF2-mediated Oxidative StressResponse 0.471 −0.392 1.3 Glutathione Redox Reactions I −0.302 −0.2772.496 Tryptophan Degradation III (Eukaryotic) 0.378 −1.265 3.742 TCACycle II (Eukaryotic) 0.277 −0.535 3.207 Glycolysis I 0.577 0 3.051Gluconeogenesis I 0.905 −0.302 3.464 Cardiac Hypertrophy Signaling(Enhanced) 0.361 −0.396 −6.236 Hepatic Fibrosis Signaling Pathway −0.25−3.181 −5.058 PDGF Signaling −0.5 −2.353 −3.528 GP6 Signaling Pathway 0−1.715 −4.938 NF-κB Signaling 0.507 −1.313 −4.636 PI3K Signaling in BLymphocytes 0.426 −1.333 −4.041 NF-κB Activation by Viruses 0 −0.816−3.9 PEDF Signaling 0 −1.043 −3.922 Sperm Motility 1.213 1.633 −2.263Sirtuin Signaling Pathway 0.849 0.62 −1.254 Xenobiotic MetabolismGeneral Signaling Pathway −0.577 −1.581 −0.152 Gα12/13 Signaling −1.279−0.707 −0.949 TNFR2 Signaling −1.89 0 −1.291 Relaxin Signaling −0.535−0.853 −2.449 Paxillin Signaling −1 −0.626 −2.268 Integrin Signaling−0.928 −1.342 −3.311 Acute Phase Response Signaling −1.177 −1.298 −2.832Small Cell Lung Cancer Signaling −0.905 −1.886 −2.236 Lymphotoxin βReceptor Signaling −1.508 −1.291 −2.183 GM-CSF Signaling −0.243 −1.46−1.225 Acute Myeloid Leukemia Signaling −0.258 −0.853 −1.134Sphingosine-1-phosphate Signaling −0.218 −0.73 −1.808 PI3K/AKT Signaling0.447 −0.18 −1.947 IGF-1 Signaling 0.243 −0.408 −1.633 GlioblastomaMultiforme Signaling 1 −0.16 −3.464 Thrombin Signaling 1.3 −0.926 −2.828Gαq Signaling 1 −0.822 −2.16 Glioma Signaling 0.5 −0.816 −2.353 IL-8Signaling 0.365 −0.905 −2.828 HGF Signaling −0.688 0 −3.182 Toll-likeReceptor Signaling −0.302 −0.258 −2.982 Macropinocytosis Signaling 0.2770.243 −2.828 LPS/IL-1 Mediated Inhibition of RXR Function 0.535 0.626−2.746 STAT3 Pathway −0.218 0.192 −2.556 CXCR4 Signaling 0 0.333 −2.214

TABLE 5 Gene Set Enrichment Analysis showing enrichment in hallmark genesets following treatment of g-ddY mice with atrasentan at 0 (control),10, 20 or 30 mg/kg/day. NES is the normalized enrichment scoreindicating accounting for gene set size. NES of <−1.5 or >1.5 wasconsidered to be biologically significant. Adj p-value is the estimatedprobability that the results of the given enriched gene set is a falsepositive finding. Atrasentan 10 Atrasentan 20 Atrasentan 30 mg/kg/dayvs. control mg/kg/day vs. control mg/kg/day vs. control Adj p- Adj p-Adj p- Pathway value NES value NES value NES HALLMARK_ADIPOGENESIS6.03E−09 2.15 3.57E−10 2.26 6.25E−10 2.64 HALLMARK_ALLOGRAFT_REJECTION1.42E−09 −2.25 3.57E−10 −2.90 2.41E−07 −2.12 HALLMARK_ANDROGEN_RESPONSE0.75 0.88 0.46 1.03 0.67 0.92 HALLMARK_ANGIOGENESIS 7.37E−04 −1.941.80E−05 −2.19 3.42E−03 −1.86 HALLMARK_APICAL_JUNCTION 1.06E−04 −1.793.57E−10 −2.24 1.23E−06 −2.00 HALLMARK_APICAL_SURFACE 5.06E−02 −1.528.79E−03 −1.67 0.03 −1.65 HALLMARK_APOPTOSIS 2.22E−03 −1.59 1.04E−08−2.17 6.12E−03 −1.52 HALLMARK_BILE_ACID_METABOLISM 6.97E−06 2.084.69E−09 2.39 1.42E−08 2.40 HALLMARK_CHOLESTEROL_HOMEOSTASIS 0.47 1.030.46 −1.03 0.27 1.14 HALLMARK_COAGULATION 1.38E−2 −1.53 4.51E−07 −2.116.12E−03 −1.56 HALLMARK_COMPLEMENT 7.80E−03 −1.51 3.81E−07 −2.013.50E−03 −1.56 HALLMARK_DNA_REPAIR 0.94 0.79 0.93 0.83 3.50E−03 1.52HALLMARK_E2F_TARGETS 3.92E−08 −2.07 3.57E−10 −2.34 1.19E−02 −1.46HALLMARK_EPITHELIAL_MESENCHYMAL_TRANSITION 8.33E−10 −2.47 3.57E−10 −2.926.25E−10 −2.60 HALLMARK_ESTROGEN_RESPONSE_EARLY 0.09 −1.27 3.84E−03−1.52 0.10 −1.22 HALLMARK_ESTROGEN_RESPONSE_LATE 0.18 −1.18 2.80E−02−1.37 0.18 1.16 HALLMARK_FATTY_ACID_METABOLISM 5.83E−09 2.25 3.57E−102.51 6.25E−10 2.90 HALLMARK_G2M_CHECKPOINT 4.73E−09 −2.15 3.57E−10 −2.264.82E−06 −1.92 HALLMARK_GLYCOLYSIS 3.80E−04 1.62 1.52E−03 1.53 1.10E−061.96 HALLMARK_HEDGEHOG_SIGNALING 5.56E−02 −1.51 8.79E−03 −1.72 1.76E−03−1.94 HALLMARK_HEME_METABOLISM 1.38E−04 1.57 4.00E−06 1.92 2.51E−07 1.99HALLMARK_HYPOXIA 2.81E−02 −1.38 1.65E−02 −1.62 0.17 −1.18HALLMARK_IL2_STAT5_SIGNALING 1.28E−04 −1.77 1.38E−09 −2.20 3.44E−03−1.56 HALLMARK_IL6_JAK_STAT3_SIGNALING 9.50E−08 −2.34 3.57E−10 −2.921.07E−07 −2.33 HALLMARK_INFLAMMATORY_RESPONSE 8.33E−10 −2.34 3.57E−10−2.96 6.25E−10 −2.40 HALLMARK_INTERFERON_ALPHA_RESPONSE 4.12E−05 −1.961.48E−09 −2.44 5.93E−04 −1.80 HALLMARK_INTERFERON_GAMMA_RESPONSE8.33E−10 −2.26 3.57E−10 −2.88 8.95E−10 −2.26 HALLMARK_KRAS_SIGNALING_DN0.10 1.28 0.08 1.32 0.26 −1.12 HALLMARK_KRAS_SIGNALING_UP 8.33E−10 −2.243.57E−10 −2.65 7.40E−10 −2.31 HALLMARK_MITOTIC_SPINDLE 3.13E−03 −1.551.61E−05 −1.79 9.54E−08 −2.02 HALLMARK_MTORC1_SIGNALING 0.28 1.11 0.111.23 2.36E−05 1.81 HALLMARK_MYC_TARGETS_V1 0.18 −1.17 0.94 −0.81 0.091.26 HALLMARK_MYC_TARGETS_V2 0.88 −0.780 0.94 0.76 0.64 0.92HALLMARK_MYOGENESIS 0.18 −1.19 2.82E−03 −1.60 0.07 −1.30HALLMARK_NOTCH_SIGNALING 0.76 0.83 0.94 0.69 0.29 −1.15HALLMARK_OXIDATIVE_PHOSPHORYLATION 8.33E−10 3.06 3.57E−10 3.38 6.25E−103.69 HALLMARK_P53_PATHWAY 0.25 −1.12 0.06 −1.3 0.27 1.10HALLMARK_PANCREAS_BETA_CELLS 0.75 0.85 0.51 0.98 0.39 1.07HALLMARK_PEROXISOME 3.55E−03 1.68 2.66E−04 1.88 6.93E−08 2.39HALLMARK_PI3K_AKT_MTOR_SIGNALING 0.73 −0.92 0.12 −1.24 0.86 0.83HALLMARK_PROTEIN_SECRETION 0.42 1.06 0.34 1.08 2.75E−03 1.73HALLMARK_REACTIVE_OXYGEN_SPECIES_PATHWAY 0.02 1.59 9.30E−04 1.616.02E−04 1.98 HALLMARK_SPERMATOGENESIS 0.74 0.89 0.21 1.17 0.20 1.21HALLMARK_TGF_BETA_SIGNALING 7.80E−03 −1.72 2.66E−04 −1.96 0.03 −1.55HALLMARK_TNFA_SIGNALING_VIA_NFKB 8.33E−10 −2.55 3.57E−10 −3.06 6.25E−10−2.63 HALLMARK_UNFOLDED_PROTEIN_RESPONSE 0.73 −0.91 0.99 −0.64 0.90 0.78HALLMARK_UV_RESPONSE_DN 1.64E−08 −2.19 3.57E−10 −2.36 6.25E−10 −2.39HALLMARK_UV_RESPONSE_UP 0.08 1.28 0.22 1.15 2.13E−03 1.63HALLMARK_WNT_BETA_CATENIN_SIGNALING 0.50 −1.02 0.13 −1.57 0.02 −1.63HALLMARK_XENOBIOTIC_METABOLISM 4.98E−07 1.95 9.17E−09 2.16 6.25E−10 2.38

TABLE 6 List of upstream regulators grouped by cytokine and growthfactor molecular types. P-value of overlap indicates the significance ofenrichment based on the number of overlapping genes between the datasetand the gene targets in IPKB. Activation z-score threshold of <−2 or >2and p-value of overlap threshold of <0.05 was considered significant.Upstream Regulator Molecule Type Predicted Activation State Activationz-score p-value of overlap VEGFB growth factor Activated 3.128 0.0183CCN5 growth factor Activated 2.45 0.000199 DKK1 growth factor Activated2.113 0.0264 GDF2 growth factor Inhibited −2.055 0.0000181 PF4 cytokineInhibited −2.219 0.0331 CCL5 cytokine Inhibited −2.268 0.00286 BMP2growth factor Inhibited −2.435 0.00524 FGF23 growth factor Inhibited−2.497 0.000088 VEGFA growth factor Inhibited −2.501  5.6E−09 TNFSF11cytokine Inhibited −2.519 0.000579 TSLP cytokine Inhibited −2.533 0.0371PDGFB growth factor Inhibited −2.622 0.0453 TGFB2 growth factorInhibited −2.631 0.0000392 TGFB3 growth factor Inhibited −2.663 0.00265IGF1 growth factor Inhibited −2.676 0.0000051 IL10 cytokine Inhibited−2.775 0.000086 CXCL12 cytokine Inhibited −2.851 0.000837 IL21 cytokineInhibited −2.886 0.0482 CSF3 cytokine Inhibited −3.116 0.000333 TNFSF12cytokine Inhibited −3.271 0.0184 IFNA2 cytokine Inhibited −3.465 0.0171NRG1 growth factor Inhibited −3.478 0.000000103 PRL cytokine Inhibited−3.529 1.13E−10 CD40LG cytokine Inhibited −3.607 0.0000006 EGF growthfactor Inhibited −3.649 7.11E−09 WNT3A cytokine Inhibited −3.7113.78E−08 IL33 cytokine Inhibited −3.781 0.00000473 IL6 cytokineInhibited −3.872 0.00000371 IL3 cytokine Inhibited −3.894 6.97E−08 IL2cytokine Inhibited −4.023 0.00000511 EDN1 cytokine Inhibited −4.260.00012 FGF2 growth factor Inhibited −4.33 3.56E−09 IFNG cytokineInhibited −4.604 1.34E−12 AGT growth factor Inhibited −4.741 5.66E−13IL1A cytokine Inhibited −4.942 0.00513 HGF growth factor Inhibited−5.138 1.15E−10 TGFB1 growth factor Inhibited −5.163 1.87E−26 CSF2cytokine Inhibited −5.645 7.08E−10 OSM cytokine Inhibited −5.9120.000000269 TNF cytokine Inhibited −6.593 5.21E−21 IL1B cytokineInhibited −7.154 4.35E−09

TABLE 7 List of upstream regulators grouped by transmembrane receptor,G-protein coupled receptor, and protein complexes molecular types.P-value of overlap indicates the significance of enrichment based on thenumber of overlapping genes between the dataset and the gene targets inIPKB. Activation z-score threshold of <−2 or >2 and p-value of overlapthreshold of <0.05 was considered significant. Upstream PredictedActivation Activation z- p-value of Regulator Molecule Type State scoreoverlap PLA2R1 transmembrane receptor Activated 2.917 0.0000325 FAStransmembrane receptor Activated 2.887 0.000000264 SMO G-protein coupledActivated 2.621 0.043 receptor IGF1R transmembrane receptor Activated2.165 0.00206 LGR5 transmembrane receptor Activated 2.137 0.000486Laminin complex Activated 2.043 0.00105 (complex) PTGER4 G-proteincoupled Activated 2.017 0.000049 receptor ADGRF5 G-protein coupledActivated 2 0.0437 receptor PROKR1 G-protein coupled Activated 2 0.0437receptor 26s Proteasome complex Inhibited −2.122 0.0000336 IL6Rtransmembrane receptor Inhibited −2.179 0.0213 FCGR1A transmembranereceptor Inhibited −2.201 0.000821 HTR7 G-protein coupled Inhibited−2.219 0.0141 receptor TCR complex Inhibited −2.362 8.88E−11 TYROBPtransmembrane receptor Inhibited −2.376 0.048 CHRM3 G-protein coupledInhibited −2.425 0.0079 receptor TNFRSF1A transmembrane receptorInhibited −2.449 0.0455 EDNRA transmembrane receptor Inhibited −2.4490.0119 Pdgf (complex) complex Inhibited −2.779 0.015 F2R G-proteincoupled Inhibited −3.04 0.00000844 receptor IFNAR1 transmembranereceptor Inhibited −3.094 0.0185 PI3K (complex) complex Inhibited −3.2930.0000027 Ifn gamma complex Inhibited −3.354 0.0476 GPER1 G-proteincoupled Inhibited −3.527 0.000000242 receptor LDL complex Inhibited−3.568 0.000272 TREM1 transmembrane receptor Inhibited −3.62 0.000494CCR2 G-protein coupled Inhibited −3.791 0.00113 receptor F3transmembrane receptor Inhibited −3.862 0.0113 PDGF BB complex Inhibited−3.976 6.87E−14 CG complex Inhibited −4.34 1.58E−10 ILR4 transmembranereceptor Inhibited −4.561 0.000392 Ige complex Inhibited −4.8830.00000228 TLR9 transmembrane receptor Inhibited −4.894 0.00697 TLR3transmembrane receptor Inhibited −5.02 0.0158 NFkB (complex) complexInhibited −5.503 0.0000354

TABLE 8 List of upstream regulators grouped by transcription regulatorand ligand-dependent nuclear receptor molecular types. P-value ofoverlap indicates the significance of enrichment based on the number ofoverlapping genes between the dataset and the gene targets in IPKB.Activation z-score threshold of <−2 or >2 and p-value of overlapthreshold of <0.05 was considered significant. Upstream PredictedActivation Activation z- p-value of Regulator Molecule Type State scoreoverlap PPARGC1A transcription regulator Activated 8.123 1.65E−14 HNF4Atranscription regulator Activated 4.856 3.88E−27 MYCN transcriptionregulator Activated 4.584 1.75E−10 RB1 transcription regulator Activated4.435 1.57E−09 SPDEF transcription regulator Activated 4.415 0.000000425LHX1 transcription regulator Activated 4.214 3.15E−15 HNF1Atranscription regulator Activated 4.107 0.0000186 PITX2 transcriptionregulator Activated 3.976 0.000000278 PPARG ligand-dependent nuclearActivated 3.581 3.16E−10 receptor XBP1 transcription regulator Activated3.505 0.00651 SMAD7 transcription regulator Activated 3.504 0.0218 IKZF1transcription regulator Activated 3.472 0.0000024 KLF15 transcriptionregulator Activated 3.401 0.000534 NFE2L2 transcription regulatorActivated 3.389 2.21E−08 RUNX3 transcription regulator Activated 3.3550.0000332 PPARA ligand-dependent nuclear Activated 3.182 3.79E−17receptor IKZF3 transcription regulator Activated 3.165 0.000634 NEUROG1transcription regulator Activated 3 0.0000441 RXRA ligand-dependentnuclear Activated 2.96 0.00000779 receptor HNF1B transcription regulatorActivated 2.944 0.0000399 TFAP2C transcription regulator Activated 2.8990.00026 MLXIPL transcription regulator Activated 2.871 0.00638 GFI1transcription regulator Activated 2.82 0.000832 KLF2 transcriptionregulator Activated 2.742 0.00107 PPARGC1B transcription regulatorActivated 2.664 0.000802 TFEB transcription regulator Activated 2.5870.000232 MEOX2 transcription regulator Activated 2.54 0.0197 ESRRAligand-dependent nuclear Activated 2.517 0.0189 receptor BCL6transcription regulator Activated 2.514 9.52E−09 GPS2 transcriptionregulator Activated 2.51 0.00975 TCF7L1 transcription regulatorActivated 2.373 0.0384 CBX5 transcription regulator Activated 2.3550.0000488 DACH1 transcription regulator Activated 2.354 0.0243 FOXO3transcription regulator Activated 2.338 9.92E−08 SRSF2 transcriptionregulator Activated 2.328 0.00808 PRDM1 transcription regulatorActivated 2.281 0.000765 SREBF2 transcription regulator Activated 2.2770.0488 SMYD1 transcription regulator Activated 2.213 1.71E−11 AHRligand-dependent nuclear Activated 2.092 3.17E−13 receptor PCGF2transcription regulator Activated 2 0.0127 Msx3 transcription regulatorInhibited −2 0.0128 SPZ1 transcription regulator Inhibited −2.022 0.0283HIF1A transcription regulator Inhibited −2.042 5.35E−08 ARNT2transcription regulator Inhibited −2.079 0.00156 ETS1 transcriptionregulator Inhibited −2.096 0.00125 TEAD4 transcription regulatorInhibited −2.143 0.00305 NCOA3 transcription regulator Inhibited −2.1430.0176 ELK1 transcription regulator Inhibited −2.151 0.00543 CEBPDtranscription regulator Inhibited −2.154 0.00211 NR3C2 ligand-dependentnuclear Inhibited −2.191 0.000655 receptor KMT2A transcription regulatorInhibited −2.246 0.000564 FOXF1 transcription regulator Inhibited −2.2530.00866 NRIP1 transcription regulator Inhibited −2.267 0.0000719 FOXM1transcription regulator Inhibited −2.307 0.000132 SMAD2 transcriptionregulator Inhibited −2.323 0.00988 HOXC6 transcription regulatorInhibited −2.35 0.000765 SOX4 transcription regulator Inhibited −2.3970.0000114 STAT4 transcription regulator Inhibited −2.401 7.22E−08 SMAD4transcription regulator Inhibited −2.417 0.00000815 MECP2 transcriptionregulator Inhibited −2.419 0.0155 ESR1 ligand-dependent nuclearInhibited −2.439 7.07E−18 receptor SP1 transcription regulator Inhibited−2.444 7.53E−10 MESP2 transcription regulator Inhibited −2.449 0.0255REL transcription regulator Inhibited −2.451 0.0000517 NR4A1ligand-dependent nuclear Inhibited −2.488 7.08E−13 receptor FOXC1transcription regulator Inhibited −2.563 0.0227 IRF8 transcriptionregulator Inhibited −2.577 0.0237 MESP1 transcription regulatorInhibited −2.646 0.0178 SNAI1 transcription regulator Inhibited −2.6996.16E−08 ECSIT transcription regulator Inhibited −2.72 0.00604 EGR1transcription regulator Inhibited −2.766 0.00543 TCF4 transcriptionregulator Inhibited −2.836 0.00000487 PAX7 transcription regulatorInhibited −2.883 0.00101 SPIB transcription regulator Inhibited −2.9330.00407 MTPN transcription regulator Inhibited −2.942 0.000223 SMAD3transcription regulator Inhibited −2.952 0.000126 PDX1 transcriptionregulator Inhibited −2.958 0.0000197 TWIST1 transcription regulatorInhibited −2.987 0.000114 CCND1 transcription regulator Inhibited −3.0031.83E−08 TEAD1 transcription regulator Inhibited −3.036 0.00192 MEF2Dtranscription regulator Inhibited −3.056 0.000802 FOXL2 transcriptionregulator Inhibited −3.108 0.00539 TEAD2 transcription regulatorInhibited −3.153 0.00127 SMARCA4 transcription regulator Inhibited−3.211 1.99E−09 NFKB1 transcription regulator Inhibited −3.239 0.0000117SRF transcription regulator Inhibited −3.269 0.000056 GATA1transcription regulator Inhibited −3.309 0.00268 FOXC2 transcriptionregulator Inhibited −3.361 0.048 TP53 transcription regulator Inhibited−3.365  1.2E−36 CTNNB1 transcription regulator Inhibited −3.373 2.83E−14SPI1 transcription regulator Inhibited −3.51 0.0135 TEAD3 transcriptionregulator Inhibited −3.638 0.0102 FEV transcription regulator Inhibited−3.651 0.000117 ERG transcription regulator Inhibited −3.672 0.0000583JUNB transcription regulator Inhibited −3.721 0.00172 MRTFAtranscription regulator Inhibited −3.84 0.0491 STAT1 transcriptionregulator Inhibited −3.918 0.00156 KLF4 transcription regulatorInhibited −3.94 0.0148 Tcf7 transcription regulator Inhibited −3.9840.0000319 YAP1 transcription regulator Inhibited −4.255 0.0000177 RELAtranscription regulator Inhibited −4.344 0.0000969 IRF7 transcriptionregulator Inhibited −4.492 0.00811 STAT3 transcription regulatorInhibited −4.716 5.94E−09 MRTFB transcription regulator Inhibited −4.7340.000158 KDM5A transcription regulator Inhibited −5.858 1.92E−13

TABLE 9 Gene expression of the components of the NF-kB signalingpathways following treatment of g-ddY mice with atrasentan at 10, 20 or30 mg/kg/day compared to atrasentan at 0 mg/kg/day (control). AtrasentanAtrasentan Atrasentan 10 mg/kg/day 20 mg/kg/day 30 mg/kg/day SymbollogFC FDR logFC FDR logFC FDR Pfkfb3 −1.16606 0.016337 −1.12478 0.009909−1.48475 0.000796 Nr4a1 −1.52471 0.02068 −1.62096 0.009287 −1.802720.001473 Gem −1.50783 0.028806 −1.23709 0.026639 −1.91184 0.002183 Ccn1−1.08749 0.035252 −1.08727 0.018504 −0.91985 0.012547 Fosl2 −0.866180.067384 −1.12215 0.016936 −1.20525 0.003639 Kiff −0.54933 0.083567−0.90396 0.009287 −0.94744 0.001843 Efna1 0.693762 0.098009 0.2860210.359139 0.727762 0.021668 F3 −0.49627 0.122022 −0.52346 0.05148 −0.5640.021548 Nfkbia −0.5525 0.123145 −0.70406 0.029887 −0.73071 0.011919Ifit2 −0.47104 0.143064 −0.63233 0.030246 −0.61195 0.016546 Snn 0.3517180.143092 0.475301 0.029887 0.472482 0.014513 Nr4a2 −1.3802 0.150927−1.11098 0.124988 −1.72799 0.022984 Klf2 −0.4894 0.156952 −0.481630.080629 −0.71822 0.011835 Jag1 −0.41987 0.163479 −0.39391 0.100075−0.67841 0.008592 Dnajb4 −0.36834 0.172404 −0.35475 0.099555 −0.753430.003454 Il1b −1.68357 0.193146 −3.02945 0.026639 −2.26303 0.029185Cdkn1a 0.558282 0.20458 0.254616 0.471505 0.636882 0.049691 Spsb1−0.61566 0.206032 −0.65404 0.094579 −1.09671 0.009718 Btg2 −0.423960.209991 −0.69738 0.029887 −0.66894 0.016296 Ccnd1 0.452977 0.2148140.643985 0.047017 0.69473 0.01924 Atf3 −1.51365 0.22778 −2.106610.060485 −2.21952 0.031678 Csf1 −0.83319 0.237302 −1.08925 0.072389−1.26006 0.027904 Trib1 −0.64809 0.240117 −0.76678 0.091587 −1.074010.019758 Zbtb10 −0.23348 0.242089 −0.25865 0.10667 −0.29122 0.049691Btg1 −0.37446 0.250264 −0.51866 0.060738 −0.48249 0.046956 Ehd1 0.2562540.260538 0.271542 0.132492 0.413845 0.022598 Rhob −0.53303 0.261164−0.54187 0.148633 −0.72645 0.045084 Nfat5 −0.31176 0.298987 −0.322990.17069 −0.58448 0.017517 Tnfaip8 0.273019 0.300195 0.399971 0.0727560.516997 0.017659 Pnrc1 0.263462 0.304842 0.041556 0.870084 0.4856270.020041 Edn1 −1.00599 0.306986 −1.32804 0.10522 −1.68836 0.035952 Rel−0.72185 0.315094 −1.0782 0.075629 −1.29814 0.026258 Nr4a3 −1.142990.340486 −1.11302 0.234364 −1.93865 0.047968 Nfkb1 −0.34242 0.36235−0.61603 0.059621 −0.64981 0.028263 Serpine1 −0.71692 0.365608 −1.501070.049438 −1.38665 0.034662 Ackr3 0.285126 0.37038 0.491654 0.0697480.548488 0.028167 Ccl20 −1.27345 0.384854 −3.56331 0.033736 −2.471080.045771 Per1 −0.41615 0.408249 −0.12155 0.798223 −0.93068 0.025478Cxcl2 −1.18965 0.446123 −2.5772 0.075629 −2.6664 0.04664 Map3k8 −0.355770.45048 −0.82217 0.050202 −0.86644 0.022718 Trai1 −0.51819 0.570487−1.48546 0.059621 −1.37067 0.046862 Id2 0.166343 0.656887 0.2227610.421533 0.539366 0.040736

TABLE 10 Expression of the components of the IL6 signaling pathwaysfollowing treatment of g-ddY mice with atrasentan at 10, 20 or 30mg/kg/day compared to atrasentan at 0 mg/kg/day (control). AtrasentanAtrasentan Atrasentan 10 mg/kg/day 20 mg/kg/day 30 mg/kg/day SymbollogFC FDR logFC FDR 20mg logFC FDR Cntfr −0.86692 0.099624 −0.857440.049966 −0.93652 0.019879 Il1b −1.68357 0.193146 −3.02945 0.026639−2.26303 0.029185 Csf1 −0.83319 0.237302 −1.08925 0.072389 −1.260060.027904 I12ra −0.85988 0.372724 −0.95904 0.206119 −1.57951 0.042633Map3k8 −0.35577 0.45048 −0.82217 0.050202 −0.86644 0.022718 Il1r1−0.29875 0.455743 −0.82629 0.029887 −0.60782 0.044115

TABLE 11 Gene expression of the components of the PDGF signalingpathways following treatment of g-ddY mice with atrasentan at 10, 20 or30 mg/kg/day compared to atrasentan at 0 mg/kg/day (control). AtrasentanAtrasentan Atrasentan 10 mg/kg/day 20 mg/kg/day 30 mg/kg/day SymbollogFC FDR logFC FDR 20mg logFC FDR Pik3r1 −0.42534 0.07447 −0.428390.033132 −0.62233 0.003454 Pdgfra −0.51003 0.113814 −0.658 0.026639−0.70937 0.008179 Nfkbia −0.5525 0.123145 −0.70406 0.029887 −0.730710.011919 Pik3cg −0.58823 0.193255 −1.09598 0.019436 −0.72874 0.034335Pla2g4a −0.69994 0.201471 −0.83808 0.067311 −0.9024 0.031577 Tiam1−0.74555 0.280074 −0.94943 0.098932 −1.22717 0.029267 Pdgfb −0.341450.351246 −0.83749 0.023432 −0.54241 0.049074 Nfkb1 −0.34242 0.36235−0.61603 0.059621 −0.64981 0.028263 Map3k1 −0.38251 0.413587 −0.720280.072782 −0.76362 0.036915 Hras 0.188685 0.470644 0.068017 0.7804010.435389 0.030337

TABLE 12 Expression of the components of the cell proliferationsignaling pathways (mitotic spindle and G2M cell cycle checkpoint)following treatment of g-ddY mice with atrasentan at 10, 20 or 30mg/kg/day compared to atrasentan at 0 mg/kg/day (control). AtrasentanAtrasentan Atrasentan 10 mg/kg/day 20 mg/kg/day 30 mg/kg/day SymbollogFC FDR logFC FDR 20 mg logFC FDR Ccdc88a −0.560213108 0.064307704−0.63737 0.019436 −0.72059 0.003971 Sorbs2 −0.395795452 0.094299303−0.47171 0.026652 −0.36011 0.033932 Rhot2 0.472719618 0.1184165950.457604 0.061542 0.543154 0.019524 Epb41l2 −0.437552327 0.123145067−0.6149 0.023432 −0.70213 0.004946 Cep192 −0.421092775 0.150917991−0.41731 0.07457 −0.43003 0.043366 Bcar1 0.337889791 0.1849619550.346027 0.092736 0.41824 0.031186 Ppp4r2 −0.421363936 0.186488374−0.39924 0.113169 −0.50412 0.036549 Tuba4a 0.273786265 0.2074462050.28609 0.100847 0.495831 0.008518 Ophn1 −0.499897642 0.209990532−0.7587 0.036699 −0.71292 0.023569 Cntrl −0.289023549 0.264775176−0.39629 0.069769 −0.55688 0.011835 Tiam1 −0.745550285 0.280074251−0.94943 0.098932 −1.22717 0.029267 Nin −0.339329706 0.291537075−0.42364 0.104354 −0.7352 0.00911 Myh10 −0.306833953 0.291537075 −0.18010.445837 −0.52418 0.023157 Flna −0.416208801 0.29963401 −0.78417 0.03471−0.61835 0.042633 Rapgef5 −0.273198499 0.310608123 −0.38035 0.087682−0.42253 0.039906 Fgd6 −0.236922692 0.318947872 −0.34192 0.082731−0.69833 0.003454 Myo1e −0.234884513 0.327254691 −0.29201 0.127987−0.41769 0.02663 Vcl −0.200128517 0.335661912 −0.33173 0.061542 −0.360450.026782 Itsn1 −0.227594667 0.384854101 −0.27517 0.181905 −0.512850.01799 Smc3 −0.230373555 0.410052693 −0.16956 0.458127 −0.4558 0.034583Myh9 −0.209349487 0.631442809 −0.45434 0.154803 −0.61714 0.04296 Slc12a2−0.250697945 0.207446205 −0.33735 0.051461 −0.33063 0.029395 Dmd−0.333454644 0.211421937 −0.34846 0.104532 −0.40498 0.043352 Ccnd10.452977372 0.214813934 0.643985 0.047017 0.69473 0.01924 Arid4a−0.277719336 0.264775176 −0.19604 0.315402 −0.57195 0.00866 Kmt5a0.260119308 0.291940817 0.287339 0.144023 0.416918 0.030303 Atrx−0.246276232 0.322185047 −0.21876 0.258831 −0.40313 0.034743 Slc38a1−0.497099916 0.335090449 −0.69888 0.099211 −0.81081 0.041274 Slc7a1−0.396018735 0.366958406 −0.73041 0.057987 −0.79523 0.02442 Ythdc1−0.193613498 0.408636337 −0.0815 0.701193 −0.35794 0.043485 Meis1−0.250703773 0.445700579 −0.35928 0.161072 −0.49344 0.045256

TABLE 13 Expression of the components of the inflammatory responsesignaling pathways following treatment of g-ddY mice with atrasentan at10, 20 or 30 mg/kg/day compared to atrasentan at 0 mg/kg/day (control).Atrasentan Atrasentan Atrasentan 10 mg/kg/day 20 mg/kg/day 30 mg/kg/daySymbol logFC FDR logFC FDR logFC FDR Rnf144b −0.71083 0.093159 −0.976490.018504 −0.88478 0.009694 Ros1 −2.07792 0.107948 −1.99181 0.057987−1.98547 0.034335 Slc7a2 −0.83149 0.120432 −1.33578 0.016936 −0.980850.018794 F3 −0.49627 0.122022 −0.52346 0.05148 −0.564 0.021548 Nfkbia−0.5525 0.123145 −0.70406 0.029887 −0.73071 0.011919 Hpn 0.4006680.134179 0.418714 0.057987 0.619746 0.006798 Slc4a4 0.512324 0.1724040.524475 0.085406 0.614581 0.03148 Chst2 −0.52123 0.191357 −0.367990.22704 −0.92143 0.008518 Il1b −1.68357 0.193146 −3.02945 0.026639−2.26303 0.029185 Cdkn1a 0.558282 0.20458 0.254616 0.471505 0.6368820.049691 Btg2 −0.42396 0.209991 −0.69738 0.029887 −0.66894 0.016296 Csf1−0.83319 0.237302 −1.08925 0.072389 −1.26006 0.027904 Slc11a2 0.3191590.264775 0.350018 0.12506 0.420512 0.049481 Eif2ak2 −0.27199 0.27558−0.30022 0.130897 −0.5033 0.015208 Edn1 −1.00599 0.306986 −1.328040.10522 −1.68836 0.035952 Nfkb1 −0.34242 0.36235 −0.61603 0.059621−0.64981 0.028263 Serpine1 −0.71692 0.365608 −1.50107 0.049438 −1.386650.034662 Slc7a1 −0.39602 0.366958 −0.73041 0.057987 −0.79523 0.02442Ccl20 −1.27345 0.384854 −3.56331 0.033736 −2.47108 0.045771 Il1r1−0.29875 0.455743 −0.82629 0.029887 −0.60782 0.044115 Ly6e 0.1666150.73244 0.243601 0.496099 0.685667 0.041805 Gabbr1 −0.02473 0.964627−0.13161 0.701936 −0.57619 0.046381

TABLE 14 Gene expression of the 44 genes associated with mesangial cellsignature following treatment of g-ddY with atrasentan at 10, 20, or 30mg/kg/day compared to atrasentan at 0 mg/kg/day (control). AtrasentanAtrasentan Atrasentan 10 mg/kg/day 20 mg/kg/day 30 mg/kg/day SymbollogFC FDR logFC FDR logFC FDR Nr4a1 −1.52471 0.02068 −1.62096 0.009287−1.80272 0.001473 Mef2c −0.62169 0.040481 −0.6302 0.019436 −0.68690.004544 Zeb2 −0.56382 0.045268 −0.64921 0.016936 −0.67777 0.003454Postn −1.38468 0.064308 −2.29303 0.009287 −1.24718 0.017778 Stard13−0.72235 0.070757 −0.63665 0.047017 −0.84655 0.008122 Rbms3 −0.574880.074375 −0.71922 0.018674 −0.88947 0.002755 Gpx3 0.680189 0.0942990.710044 0.040407 1.122196 0.003454 Ahnak −0.57555 0.103546 −0.712820.026639 −1.01336 0.002755 Nfkbia −0.5525 0.123145 −0.70406 0.029887−0.73071 0.011919 Epb4112 −0.43755 0.123145 −0.6149 0.023432 −0.702130.004946 Gata3 −0.48215 0.141854 −0.49245 0.06467 −0.52161 0.031516 Akt3−0.38631 0.143064 −0.46726 0.041092 −0.48434 0.018794 Cald1 −0.371930.159141 −0.53608 0.028056 −0.50596 0.015786 Unc5c 0.561795 0.1601940.36849 0.224278 0.951658 0.00669 Psd3 −0.35444 0.172404 −0.522730.029887 −0.58428 0.00866 Pparg −0.61292 0.172404 −0.56156 0.112215−0.67808 0.042887 Robo1 −0.48025 0.180027 −0.7261 0.029888 −0.600870.029099 C7 −0.77671 0.201471 −1.277 0.031245 −1.07912 0.026221 Raph1−0.29653 0.202275 −0.38519 0.054865 −0.53579 0.008122 Lama2 −0.295530.206144 −0.4437 0.035116 −0.48948 0.011923 Tjp1 −0.24892 0.242617−0.30541 0.081209 −0.39891 0.019552 Utm −0.49005 0.247838 −0.514340.126507 −0.76971 0.022718 Aldob 0.419369 0.259564 0.492327 0.1020040.747988 0.015448 Fchsd2 −0.25643 0.262979 −0.18823 0.292954 −0.377220.032457 Nbea −0.28187 0.270291 −0.19334 0.334672 −0.43202 0.029267 Ebf1−0.33006 0.274512 −0.35124 0.142697 −0.45065 0.047835 Tcf4 −0.217580.286673 −0.22405 0.162797 −0.30548 0.046576 Ppp1r12a −0.19949 0.294247−0.21354 0.15604 −0.38255 0.015257 Ankrd12 −0.28245 0.298987 −0.246280.245564 −0.4726 0.026737 Ptprm −0.26142 0.298987 −0.2678 0.175547−0.54534 0.011636 Ets1 −0.21985 0.308571 −0.30649 0.086114 −0.337870.040012 Rapgef5 −0.2732 0.310608 −0.38035 0.087682 −0.42253 0.039906Myo1e −0.23488 0.327255 −0.29201 0.127987 −0.41769 0.02663 Tns3 −0.26380.340113 −0.30431 0.162366 −0.45216 0.033592 Tacc1 −0.24487 0.354586−0.3693 0.090837 −0.49245 0.020907 Piezo2 −0.40781 0.391686 −0.614790.113663 −0.8237 0.030578 Maml2 −0.23179 0.42616 −0.23109 0.309113−0.44408 0.043294 Akap12 −0.2687 0.445264 −0.54041 0.068209 −0.82110.008226 Meis1 −0.2507 0.445701 −0.35928 0.161072 −0.49344 0.045256Frmd4a −0.28382 0.446123 −0.39548 0.171443 −0.57422 0.041274 Igfbp70.205172 0.45523 0.108837 0.648747 0.425852 0.040662 Prkg1 −0.218710.488645 −0.32796 0.180294 −0.63558 0.015208 Myh9 −0.20935 0.631443−0.45434 0.154803 −0.61714 0.04296 Cblb −0.082 0.788712 −0.206820.316455 −0.42884 0.034058

CONCLUSION

Thus, in the g-ddY mouse model of spontaneous IgA nephropathy,atrasentan treatment at 20 and 30 mg/kg/day significantly reducedalbuminuria following approximately 5-days of treatment compared tocontrol g-ddY mice not treated with atrasentan. The magnitude ofalbuminuria lowering observed at these doses (>60% reduction frombaseline) is considered highly clinically meaningful, demonstrating thatatrasentan reduces albuminuria in a mouse model of IgA nephropathy.

What is claimed:
 1. A method of treating IgA nephropathy in a subject inneed thereof, comprising: (a) determining that the subject has IgAnephropathy; and (b) administering to the subject a therapeuticallyeffective amount of atrasentan, or a pharmaceutically acceptable saltthereof.
 2. The method of claim 1, wherein step (a) comprises performinga kidney biopsy.
 3. The method of claim 1, wherein step (a) comprisesdetecting deposition of IgA-immune complexes in the kidney.
 4. Themethod of claim 1, wherein step (a) comprises detecting anti-glycanantibodies.
 5. The method of claim 1, wherein step (a) comprises two orthree of: performing a kidney biopsy, detecting deposition of IgA-immunecomplexes in the kidney, and detecting anti-glycan antibodies.
 6. Themethod of claim 1, wherein the therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof, is from about0.20 mg to about 1.5 mg of atrasentan, or an equivalent amount of apharmaceutically acceptable salt thereof.
 7. The method of claim 6,wherein the therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof, is from about 0.25 mg to about1.25 mg of atrasentan, or an equivalent amount of a pharmaceuticallyacceptable salt thereof.
 8. The method of claim 7, wherein thetherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, is from about 0.40 mg to about 0.85 mg ofatrasentan, or an equivalent amount of a pharmaceutically acceptablesalt thereof.
 9. The method of claim 8, wherein the therapeuticallyeffective amount of atrasentan, or a pharmaceutically acceptable saltthereof, is about 0.75 mg of atrasentan, or an equivalent amount of apharmaceutically acceptable salt thereof.
 10. The method of claim 1,wherein the atrasentan is administered as a pharmaceutically acceptablesalt.
 11. The method of claim 10, wherein the atrasentan is administeredas atrasentan hydrochloride or atrasentan mandelate.
 12. The method ofclaim 11, wherein the atrasentan is administered as atrasentanhydrochloride.
 13. The method of claim 11, wherein the atrasentan isadministered as atrasentan mandelate.
 14. The method of claim 1, whereinthe atrasentan is administered as the free base.
 15. The method of claim2, wherein the therapeutically effective amount of atrasentan, or apharmaceutically acceptable salt thereof, is from about 0.20 mg to about1.5 mg of atrasentan, or an equivalent amount of a pharmaceuticallyacceptable salt thereof.
 16. The method of claim 15, wherein thetherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, is from about 0.25 mg to about 1.25 mg ofatrasentan, or an equivalent amount of a pharmaceutically acceptablesalt thereof.
 17. The method of claim 16, wherein the therapeuticallyeffective amount of atrasentan, or a pharmaceutically acceptable saltthereof, is from about 0.40 mg to about 0.85 mg of atrasentan, or anequivalent amount of a pharmaceutically acceptable salt thereof.
 18. Themethod of claim 17, wherein the therapeutically effective amount ofatrasentan, or a pharmaceutically acceptable salt thereof, is about 0.75mg of atrasentan, or an equivalent amount of a pharmaceuticallyacceptable salt thereof.
 19. The method of claim 2, wherein theatrasentan is administered as a pharmaceutically acceptable salt. 20.The method of claim 19, wherein the atrasentan is administered asatrasentan hydrochloride or atrasentan mandelate.
 21. The method ofclaim 20, wherein the atrasentan is administered as atrasentanhydrochloride.
 22. The method of claim 20, wherein the atrasentan isadministered as atrasentan mandelate.
 23. The method of claim 2, whereinthe atrasentan is administered as the free base.
 24. The method of claim1, wherein the subject has been determined not to suffer from one ormore of diabetic nephropathy, HIV/AIDS, HIV-related nephropathy,prostate cancer, or acute kidney failure.
 25. The method of claim 2,wherein the subject has been determined not to suffer from one or moreof diabetic nephropathy, HIV-related nephropathy, prostate cancer, oracute kidney failure.
 26. The method of claim 24, wherein thetherapeutically effective amount of atrasentan, or a pharmaceuticallyacceptable salt thereof, is from about 0.20 mg to about 1.5 mg ofatrasentan, or an equivalent amount of a pharmaceutically acceptablesalt thereof.
 27. The method of claim 25, wherein the therapeuticallyeffective amount of atrasentan, or a pharmaceutically acceptable saltthereof, is from about 0.20 mg to about 1.5 mg of atrasentan, or anequivalent amount of a pharmaceutically acceptable salt thereof.
 28. Themethod of claim 26, wherein the atrasentan is administered as atrasentanhydrochloride or atrasentan mandelate.
 29. The method of claim 27,wherein the atrasentan is administered as atrasentan hydrochloride oratrasentan mandelate.